Bile acid recycling inhibitors and satiogens for treatment of diabetes, obesity, and inflammatory gastrointestinal conditions

ABSTRACT

Provided herein are methods of utilizing bile acid transport inhibitors and/or enteroendocrine peptide enhancing agents for the treatment of obesity, diabetes, and inflammatory gastrointestinal conditions.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.14/555,290, filed Nov. 26, 2014, which is a continuation of U.S.application Ser. No. 13/116,988, filed on May 26, 2011, now Abandoned,which claims the benefit under 35 U.S.C. § 119(e) of U.S. ProvisionalApplication No. 61/348,666, filed May 26, 2010, and U.S. ProvisionalApplication No. 61/348,669, filed May 26, 2010, each of which isincorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION

Diabetes and obesity affect numerous humans throughout the world, andare associated with or induces other diseases or conditions. Inparticular, diabetes and obesity are serious risk factors for diseasesand conditions such as hypertension, gallbladder disease, cancer,polycystic ovary disease and arteriosclerosis and can contribute toelevated levels of cholesterol in the blood. Overeating and obesity,which frequently leads to diabetes, have become a problem in the generalpopulation. Consequently there is interest in reducing food intake,losing weight, and reducing elevated blood glucose.

Inflammatory gastrointestinal conditions affect millions of people. Forexample, necrotizing enterocolitis (NEC) affects thousands of newbornseach year and is the most common gastrointestinal emergency of prematureinfants. Mortality rates of NEC are between 10 to 50%, and thus NECremains a major cause of morbidity and mortality in premature infants.The pathophysiology of NEC is poorly understood. An effective treatmentof NEC as well as other inflammatory gastrointestinal conditions isneeded.

SUMMARY OF THE INVENTION

Provided herein, in certain embodiments, are therapeutic methods usingcompounds that inhibit the Apical Sodium-dependent Bile Transporter(ASBT) or any recuperative bile salt transporter. In certain instances,use of the compounds provided herein reduces or inhibits recycling ofbile acid salts in the gastrointestinal tract. In some embodiments, themethods provided herein reduce intraenterocyte bile acids or reducenecrosis and/or damage to ileal architecture. In some embodiments, thebile transport inhibitors are non-systemic compounds. In otherembodiments, the bile acid transporter inhibitors are systemiccompounds. In certain embodiments, the bile transport inhibitorsdescribed herein enhance L-cell secretion of enteroendocrine peptides.

In certain embodiments, provided herein are methods for treating orpreventing an inflammatory intestinal condition or a metabolic disordercomprising administering a therapeutically effective amount of an ASBTinhibitor (ASBTI) and/or an enteroendocrine peptide enhancing agentand/or a nuclear farnesoid X receptor (FXR) agonist to an individual inneed thereof. In certain embodiments, provided herein is an ASBTI and/oran enteroendocrine peptide enhancing agent and/or a FXR agonist for usein the treatment of an inflammatory intestinal condition comprisingadministering a therapeutically effective amount of an ASBTI and/or anenteroendocrine peptide enhancing agent and/or a FXR agonist to anindividual in need thereof. In some embodiments, an ASBTI and/or anenteroendocrine peptide enhancing agent and/or a FXR agonist isminimally absorbed. In some embodiments, an ASBTI and/or anenteroendocrine peptide enhancing agent and/or a FXR agonist isnon-systemically administered to the distal ileum of an individual inneed thereof. In some embodiments, an ASBTI and/or an enteroendocrinepeptide enhancing agent and/or a FXR agonist is non-systemicallyadministered to the colon or the rectum of an individual in needthereof. In some embodiments, less than 50%, less than 40%, less than30%, less than 20%, less than 10%, less than 9%, less than 8%, less than7%, less than 6%, less than 5%, less than 4%, less than 3%, less than2%, or less than 1% of the ASBTI and/or the enteroendocrine peptideenhancing agent and/or a FXR agonist is systemically absorbed.

In some embodiments, the methods provided herein treat inflammatoryintestinal conditions including but not limited to necrotizingenterocolitis, gastritis, ulcerative colitis, inflammatory boweldisease, irritable bowel syndrome, gastroenteritis, radiation inducedenteritis, chemotherapy induced enteritis, gastro-esophageal refluxdisease (GERD), peptic ulcer, non-ulcer dyspepsia (NUD), celiac disease,intestinal celiac disease, gastrointestinal complications followingbariatric surgery, gastric carcinogenesis, or gastric carcinogenesisfollowing gastric or bowel resection.

In some embodiments, the individual is a newborn or prematurely borninfant. In some embodiments, the individual is enterally-fed infant orformula-fed infant.

In some embodiments, the methods described herein reduce intraenterocytebile acids in an individual in need thereof. In some embodiments, themethods described herein reduce accumulation of bile acids in ilealenterocytes of an individual in need thereof. In some embodiments, themethods described herein inhibit transport of bile acids from ileallumen into enterocytes of an individual in need thereof. In someembodiments, the methods described herein increase ileal luminal bileacids of an individual in need thereof. In some embodiments, the methodsdescribed herein reduce necrosis or damage to ileal architecture orileal cells in an individual in need thereof.

In some embodiments of the methods described above, the ASBTI is acompound of Formula I as described herein. In some embodiments of themethods described above, the ASBTI is a compound of Formula II asdescribed herein. In some embodiments of the methods described above,the ASBTI is a compound of Formula III as described herein. In someembodiments of the methods described above, the ASBTI is a compound ofFormula IV as described herein. In some embodiments of the methodsdescribed above, the ASBTI is a compound of Formula V as describedherein. In some embodiments of the methods described above, the ASBTI isa compound of Formula VI or Formula VID as described herein.

In certain embodiments, an ASBTI is any compound described herein thatinhibits recycling of bile acids in the gastrointestinal tract of anindividual. In certain embodiments, an ASBTI is(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxide;(“Compound 100A”) or any other salt or analog thereof. In certain of anyof the aforementioned embodiments, an ASBTI is1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate salt (“Compound 100B”) or any other salt or analogthereof. In certain embodiments, an ASBTI is N,N-dimethylimido-dicarbonimidic diamide (“Compound 100C”) or any salt oranalog thereof. In certain embodiments, an ASBTI is any commerciallyavailable ASBTI including but not limited to SD-5613, A-3309, 264W94,S-8921, SAR-548304, BARI-1741, HMR-1453, TA-7552, R-146224, or SC-435.In some embodiments, an ASBTI is1,1-Dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N—[(R)-α-[N-(2-sulphoethyl)carbamoyl]-4-hydroxybenzyl]carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;1,1-Dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N—[(R)-α-[N—((S)-1-carboxy-2-(R)-hydroxypropyl)carbamoyl]-4-hydroxybenzyl]carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;1,1-Dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N—[(R)-α-[N—((S)-1-carboxy-2-methylpropyl)carbamoyl]-4-hydroxybenzyl]carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;1-[[5-[[3-[(3S,4R,5R)-3-butyl-7-(dimethylamino)-3-ethyl-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5yl]phenyl]amino]-5-oxopentyl]amino]-1-deoxy-D-glucitol;orPotassium((2R,3R,4S,5R,6R)-4-benzyloxy-6-{3-[3-((3S,4R,5R)-3-butyl-7-dimethylamino-3-ethyl-4-hydroxy-1,1-dioxo-2,3,4,5-tetrahydro-1H-benzo[b]thiepin-5-yl)-phenyl]-ureido}-3,5-dihydroxy-tetrahydro-pyran-2-ylmethyl)sulphateethanolate, hydrate.

In certain embodiments, an enteroendocrine peptide secretion enhancingagent is a bile acid, a bile salt, a bile acid mimic, a bile salt mimic,TGR5 agonist, or a combination thereof. In some embodiments, theenteroendocrine peptide secretion enhancing agent is a glucagon-likepeptide secretion enhancing agent, optionally in combination with a bileacid, a bile salt, a bile acid mimic, or a bile salt mimic. In certainembodiments, the glucagon-like peptide secretion enhancing agent is aglucagon-like peptide-1 (GLP-1) secretion enhancing agent or aglucagon-like peptide-2 (GLP-2) secretion enhancing agent, optionally incombination with a bile acid, a bile salt, a bile acid mimic, or a bilesalt mimic. In some embodiments, the enteroendocrine peptide secretionenhancing agent is a pancreatic polypeptide-fold peptide secretionenhancing agent, optionally in combination with a bile acid, a bilesalt, a bile acid mimic, or a bile salt mimic. In some embodiments, thepancreatic polypeptide-fold peptide secretion enhancing agent is apeptide YY (PYY) secretion enhancing agent.

In certain embodiments, the FXR agonist is GW4064, GW9662, INT-747,T0901317, WAY-362450, fexaramine, a cholic acid, a deoxycholic acid, aglycocholic acid, a glycodeoxycholic acid, a taurocholic acid, ataurodihydrofusidate, a taurodeoxycholic acid, a cholate, aglycocholate, a deoxycholate, a taurocholate, a taurodeoxycholate, achenodeoxycholic acid, or a salt thereof, or a combination thereof.

Provided in certain embodiments herein are methods and dosage forms(e.g., oral or rectal dosage form) for use in the treatment of diabetes,obesity, or an inflammatory intestinal condition comprising atherapeutically effective amount of a bile acid, bile salt, or mimeticthereof, and a carrier. In some embodiments, the bile acid, bile salt,or mimetic thereof is a minimally absorbed bile acid, bile salt, ormimetic thereof. In specific embodiments, the dosage form is an entericformulation, an ileal-pH sensitive release, or a suppository or othersuitable form.

In some embodiments, provided herein is a method for treating diabetes,obesity, or an inflammatory intestinal condition comprising rectallyadministering a therapeutically effective amount of a minimally absorbedbile acid, bile salt, or mimetic thereof.

In some embodiments, the methods provided herein further compriseadministering a therapeutically effective amount of an inhibitor ofDipeptidyl Peptidase-4. In some embodiments, the inhibitor of DipeptidylPeptidase-4 is administered orally or rectally. In some embodiments, theinhibitor of Dipeptidyl Peptidase-4 is co-administered with an ASBTI, anenteroendocrine peptide enhancing agent, a FXR agonist, bile acid, bilesalt, or mimetic thereof. In some embodiments, the inhibitor ofDipeptidyl Peptidase-4 is an absorbable or systemically absorbedinhibitor of Dipeptidyl Peptidase-4.

In certain embodiments, a bile acid mimetic is a TGR5 agonist, M-BARagonist, GPR119 agonist, GPR120 agonist, GPR131 agonist, GPR140 agonist,GPR143 agonist, GPBAR1 agonist, BG37 agonist, farnesoid-X receptoragonist. In some instances, a bile-acid mimetic promotes L-cellsecretions. In certain instances, a bile acid mimetic promotes thesecretion of GLP-1, GLP-2, PYY, OXM, or a combination thereof.

In some embodiments, the methods described above further compriseadministration of a second agent selected from a liver receptor homolog1 (LRH-1), a DPP-IV inhibitor, a proton pump inhibitor, H2 antagonist,prokinetic agent, a biguanide, an incretin mimetic, a thiazolidinone, amucoadhesive agent, and GLP-1 or an analog thereof, and a TGR5 agonist.In some embodiments, the second agent is a DPP-IV inhibitor.

In some embodiments, the composition administered comprises at least oneof a spreading agent or a wetting agent. In some embodiments, theabsorption inhibitor is a mucoadhesive agent (e.g., a mucoadhesivepolymer). In certain embodiments, the mucoadhesive agent is selectedfrom methyl cellulose, polycarbophil, polyvinylpyrrolidone, sodiumcarboxymethyl cellulose, and combinations thereof. In some embodiments,a pharmaceutical composition administered further comprises anenteroendocrine peptide.

In some embodiments, provided herein is a method for lowering elevatedblood glucose in a mammal resulting from food intake comprising orallyadministering to said mammal a therapeutically effective amount of aminimally absorbed Apical Sodium-dependent Bile Transporter inhibitor.In some embodiments, provided herein is a method further comprisingadministering to said mammal a therapeutically effective amount of aninhibitor of Dipeptidyl Peptidase-4.

In specific instances, provided herein is a method for lowering elevatedbody weight or lowering elevated blood glucose in a mammal resultingfrom food intake comprising orally administering to said mammal atherapeutically effective amount of a minimally absorbed ApicalSodium-dependent Bile Transporter inhibitor and/or an inhibitor ofDipeptidyl Peptidase-4. In other embodiments, provided herein is amethod for lowering elevated body weight or lowering elevated bloodglucose in a mammal resulting from food intake comprising orallyco-administering to said mammal a therapeutically effective amount of aminimally absorbed Apical Sodium-dependent Bile Transporter inhibitorand an inhibitor of Dipeptidyl Peptidase-4.

Also provided herein is a method of promoting stimulation of L-cellsecretion in an individual in need thereof, the method comprising orallyor rectally administering an effective amount of a minimally absorbedbile acid, bile salt, or mimetic thereof. In specific instances, theindividual in need thereof is suffering from a disorder ameliorated byL-cell secreted products.

In certain embodiments, increased L-cell secretion of enteroendocrinepeptides is associated with reduced necrosis and/or reduced damage toileal architecture. In certain instances, increased L-cell secretion ofenteroendocrine peptides is associated with induction of satiety and/orreduction of food intake (caloric intake) and subsequent weight loss. Insome embodiments, increased L-cell secretion of enteroendocrine peptidesis associated with a reduction in blood and/or plasma glucose levels ina hyperglycemic individual. In some instances, increased L-cellsecretion of enteroendocrine peptides is associated with increasedinsulin sensitivity.

In some embodiments, provided herein is a pharmaceutical compositionformulated for non-systemic ileal, rectal or colonic delivery of theenteroendocrine peptide secretion enhancing agent. In certainembodiments, provided herein is an enteroendocrine peptide secretionenhancing agent such as a TGR5 agonist, a bile acid, a bile salt, a bileacid mimic, a bile salt mimic, or a combination thereof. In someembodiments, the enteroendocrine peptide secretion enhancing agent is aglucagon-like peptide secretion enhancing agent, optionally incombination with a bile acid, a bile salt, a bile acid mimic, or a bilesalt mimic. In certain embodiments, the glucagon-like peptide secretionenhancing agent is a glucagon-like peptide-1 (GLP-1) secretion enhancingagent or a glucagon-like peptide-2 (GLP-2) secretion enhancing agent,optionally in combination with a bile acid, a bile salt, a bile acidmimic, or a bile salt mimic. In some embodiments, the enteroendocrinepeptide secretion enhancing agent is a pancreatic polypeptide-foldpeptide secretion enhancing agent, optionally in combination with a bileacid, a bile salt, a bile acid mimic, or a bile salt mimic. In someembodiments, the pancreatic polypeptide-fold peptide secretion enhancingagent is a peptide YY (PYY) secretion enhancing agent.

In certain embodiments, the composition further comprises at least oneof a cholesterol absorption inhibitor, a spreading agent or a wettingagent. In some embodiments, the absorption inhibitor is a mucoadhesiveagent. In specific embodiments, the polymer having mucoadhesiveproperties is selected from methyl cellulose, polycarbophil,polyvinylpyrrolidone, sodium carboxymethyl cellulose, and combinationsthereof. In some embodiments, the enteroendocrine peptide secretionenhancing agent is covalently linked to the absorption inhibitor.

In certain embodiments, the carrier is a rectally suitable carrier. Incertain embodiments, any pharmaceutical composition described herein isformulated as a suppository, an enema solution, a rectal foam, or arectal gel. In some embodiments, any pharmaceutical compositiondescribed herein comprises an orally suitable carrier. In certainembodiments, the pharmaceutical composition comprises an entericcoating.

Provided in certain embodiments herein is a method for treating ametabolic disease or a condition associated with a metabolic diseasecomprising administering to the lower ileum, the colon and/or the rectumof an individual in need thereof, a pharmaceutical compositioncomprising a therapeutically effective amount of an enteroendocrinepeptide secretion enhancing agent. Provided in certain embodimentsherein is a method for treating a metabolic disease or a conditionassociated with a metabolic disease or an inflammatory intestinalcondition comprising administering (e.g., orally or rectallyadministering) to an individual in need thereof, a pharmaceuticalcomposition comprising a therapeutically effective amount of anenteroendocrine peptide secretion enhancing agent and a carrier. In someembodiments, compositions provided herein comprise administering atherapeutically effective amount of ASBTI. In some embodiments,administered is a pharmaceutical composition that further comprises anabsorption inhibitor, wherein the absorption inhibitor inhibits theabsorption of the enteroendocrine peptide secretion enhancing agentacross the rectal or colonic mucosa. In some embodiments, thecomposition administered comprises an orally suitable carrier. Incertain embodiments, the pharmaceutical composition is formulated forenteric delivery. In some embodiments, the pharmaceutical compositioncomprises an enteric coating.

Provided in certain embodiments herein is a method for treating obesity,diabetes, or an inflammatory intestinal condition comprisingadministering to the lower ileum, the colon and/or the rectum of anindividual in need thereof, a pharmaceutical composition comprising atherapeutically effective amount of an enteroendocrine peptide secretionenhancing agent. Provided in some embodiments herein is a method fortreating obesity, diabetes, or an inflammatory intestinal conditioncomprising administering (e.g., orally or rectally administering) to anindividual in need thereof, a pharmaceutical composition comprising atherapeutically effective amount of an enteroendocrine peptide secretionenhancing agent and a carrier. In some embodiments, compositionsprovided herein further comprise administering a therapeuticallyeffective amount of ASBTI. In some embodiments, administered is apharmaceutical composition that further comprises an absorptioninhibitor, wherein the absorption inhibitor inhibits the absorption ofthe enteroendocrine peptide secretion enhancing agent across the rectalor colonic mucosa. In some embodiments, the composition administeredcomprises an orally suitable carrier. In certain embodiments, thepharmaceutical composition is formulated for enteric delivery. In someembodiments, the pharmaceutical composition comprises an entericcoating.

In certain other embodiments, provided herein is a method of treating(e.g., preventing, prophylactically treating or reducing the incidencesof) pancreatic and other cancers comprising contacting the distal ileumof an individual in need thereof with an Apical Sodium-dependent BileTransporter inhibitor.

Provided in certain embodiments herein is a method for preventing ortreating pancreatic cancers comprising administering to the lower ileum,the colon and/or the rectum of an individual in need thereof, apharmaceutical composition comprising a therapeutically effective amountof an enteroendocrine peptide secretion enhancing agent. Provided insome embodiments herein is a method for preventing or treatingpancreatic cancers comprising administering (e.g., orally or rectallyadministering) to an individual in need thereof, a pharmaceuticalcomposition comprising a therapeutically effective amount of anenteroendocrine peptide secretion enhancing agent and a carrier. In someembodiments, administered is a pharmaceutical composition that furthercomprises an absorption inhibitor, wherein the absorption inhibitorinhibits the absorption of the enteroendocrine peptide secretionenhancing agent across the rectal or colonic mucosa. In someembodiments, the composition administered comprises an orally suitablecarrier. In certain embodiments, the pharmaceutical composition isformulated for enteric delivery. In some embodiments, the pharmaceuticalcomposition comprises an enteric coating.

In some embodiments, administered according to a method described hereinis a composition comprising a rectally suitable carrier. In someembodiments, the administered pharmaceutical composition is formulatedas a suppository, an enema solution, a rectal foam, or a rectal gel.

In certain embodiments, a composition administered according to a methoddescribed herein comprises an enteroendocrine peptide secretionenhancing agent that is a bile acid, bile salt, bile acid mimic or bilesalt mimic. In some embodiments, the enteroendocrine peptide secretionenhancing agent is a glucagon-like peptide secretion enhancing agent. Incertain embodiments, the enteroendocrine peptide secretion enhancingagent is a glucagon-like peptide-1 (GLP-1) secretion enhancing agent ora glucagon-like peptide-2 (GLP-2) secretion enhancing agent. In someembodiments, the enteroendocrine peptide secretion enhancing agent is apancreatic polypeptide-fold peptide secretion enhancing agent. Inspecific embodiments, the enteroendocrine peptide secretion enhancingagent is a peptide YY (PYY) secretion enhancing agent. In someembodiments, a composition administered according to a method describedherein comprises any one or more of the enteroendocrine peptidesecretion enhancing agents described herein.

Without limiting their utility to appetite suppression, agents deliveredrectally that invoke enteroendocrine signals will be termed, forconvenience, “rectal satiogens”. As an example of how this definitionshould not be limiting, the stimulation of enteroendocrine regulatorypeptides can benefit not only metabolic diseases by the above effects,but can benefit a number of other conditions via mechanisms that may beentirely independent of the control of nutrient flux. Examples ofconditions in which enhanced enteroendocrine secretion can be beneficialare described in US 2009/0264808, and include: diabetes, impairedglucose tolerance, glucose metabolic disorders, insulin resistance,obesity, acute coronary syndrome, hibernating myocardium, ventriculardysfunction, cardiac risk, post myocardial infarction mortality,post-surgical or sepsis-related or critical illness-related catabolismand mortality, critical illness polyneuropathy, congestive heartfailure, toxic hypervolemia, renal failure, ischemia-reperfusion injury,mortality and morbidity from stroke and neurodegenerative disease,neuropathy, inflammatory intestinal conditions, inflamatory boweldisease, necrotizing enterocolitis, gastritis, ulcerative colitis,inflammatory bowel disease, irritable bowel syndrome, gastroenteritis,radiation induced enteritis, chemotherapy induced enteritis,gastro-esophageal reflux disease (GERD), peptic ulcer, non-ulcerdyspepsia (NUD), celiac disease, intestinal celiac disease,gastrointestinal complications following bariatric surgery, gastriccarcinogenesis, or gastric carcinogenesis following gastric or bowelresection, bowel mucosal injury, impaired bowel integrity, osteopenia,and bone fractures and bone disorders.

Provided herein are methods and uses for treating obesity or diabetes oran inflammatory intestinal condition comprising contacting the distalileum of an individual in need thereof with an Apical Sodium-dependentBile Transporter Inhibitor (ASBTI) and/or enteroendocrine peptideenhancing agent and/or a FXR agonist.

In some embodiments of the methods and uses, contacting the distal ileumof an individual in need thereof with an ASBTI and/or enteroendocrinepeptide enhancing agent and/or a FXR agonist:

-   -   a. reduces intraenterocyte bile acids;    -   b. reduces necrosis and/or damage to ileal architecture;    -   c. reduces food intake in the individual;    -   d. induces satiety in the individual;    -   e. reduces blood and/or plasma glucose levels in the individual;    -   f. treats a metabolic disorder in the individual;    -   g. reduces the weight of the individual;    -   h. stimulates L-cells in the distal gastrointestinal tract of        the individual;    -   i. increases the concentration of bile acids and salts thereof        in the vicinity of L-cells in the distal gastrointestinal tract        of the individual;    -   j. enhances enteroendocrine peptide secretion in the individual;        or    -   k. any combination thereof.

In some embodiments, the ASBTI and/or the enteroendocrine peptideenhancing agent and/or a FXR agonist is not systemically absorbed. Insome other embodiments, the ASBTI and/or the enteroendocrine peptideenhancing agent and/or a FXR agonist is systemically absorbed.

In some embodiments of the methods, the individual is an obese ormorbidly overweight individual. In some embodiments of the methods, theindividual is a diabetic individual. In some embodiments of the methods,the individual is a non-diabetic individual.

In some embodiments, provided herein are methods for the treatment ofobesity, diabetes, or an inflammatory intestinal condition comprisingadministration of a therapeutically effective amount of a combination ofan ASBTI and a DPP-IV inhibitor to an individual in need thereof. Insome embodiments, provided herein are methods for the treatment ofobesity, diabetes, or an inflammatory intestinal condition comprisingadministration of a therapeutically effective amount of a combination ofan ASBTI and an enteroendocrine peptide enhancing agent and/or a FXRagonist to an individual in need thereof. In some embodiments, providedherein are methods for the treatment of obesity, diabetes, or aninflammatory intestinal condition comprising administration of atherapeutically effective amount of a combination of an ASBTI and athiazolidinedione to an individual in need thereof. In some embodiments,provided herein are methods for the treatment of obesity, diabetes, oran inflammatory intestinal condition comprising administration of atherapeutically effective amount of a combination of an ASBTI and anincretin mimic to an individual in need thereof. In some embodiments,provided herein are methods for the treatment of obesity, diabetes, oran inflammatory intestinal condition comprising administration of atherapeutically effective amount of a combination of an ASBTI and GLP-1or an analog thereof to an individual in need thereof. In someembodiments, provided herein are methods for the treatment of obesity,diabetes, or an inflammatory intestinal condition comprisingadministration of a therapeutically effective amount of a combination ofan ASBTI and a biguanide to an individual in need thereof.

In some embodiments, the methods described herein reduce intraenterocytebile acids in an individual in need thereof. In some embodiments, themethods described herein reduce necrosis or damage to ileal architecturein an individual in need thereof. In some embodiments, the methodsdescribed herein reduce food intake (caloric intake) in an individual inneed thereof. In some embodiments, the methods described herein inducesatiety in an individual in need thereof. In some embodiments, themethods described herein treat metabolic disorders in an individual inneed thereof. In some embodiments, the methods described herein reducethe weight of an individual in need thereof. In some embodiments, themethods described herein stimulate L-cells in the distalgastrointestinal tract of an individual in need thereof. In someembodiments, the methods described herein increase the concentration ofbile acid and salts thereof in the vicinity of L-cells in the distalgastrointestinal tract of an individual.

Provided herein are methods for reducing food intake in an individual inneed thereof comprising administration of an Apical Sodium-dependentBile Acid Transporter Inhibitor (ASBTI) to an individual in need thereofwherein the ASBTI is delivered or released non-systemically in thedistal ileum of the individual.

Provided herein are methods for reducing circulating blood or plasmaglucose levels in an individual in need thereof comprisingadministration of an Apical Sodium-dependent Bile Acid TransporterInhibitor (ASBTI) to an individual in need thereof wherein the ASBTI isdelivered or released non-systemically in the distal ileum of theindividual.

Provided herein are methods for increasing insulin secretion in anindividual in need thereof comprising administration of an ApicalSodium-dependent Bile Acid Transporter Inhibitor (ASBTI) to anindividual in need thereof wherein the ASBTI is delivered or releasednon-systemically in the distal ileum of the individual.

In some embodiments, the methods described herein enhanceenteroendocrine peptide secretion in an individual in need thereof. Insome of such embodiments, the enteroendocrine peptide is GLP-1, GLP-2,PYY, oxyntomodulin, or a combination thereof.

In some embodiments, contacting the distal ileum of an individual inneed thereof with an ASBTI and/or an enterendocrine peptide enhancingagent and/or a FXR agonist increases the level of GLP-1 in the bloodand/or plasma of the individual by from about 2 times to about 6 timesthe level of GLP-1 in the blood and/or plasma of the individual prior tocontacting the distal ileum of the individual with the ASBTI.

In some embodiments, contacting the distal ileum of an individual inneed thereof with an ASBTI reduces the level of glucose in the bloodand/or plasma of the individual by at least 30% compared to the level ofglucose in the blood and/or plasma of the individual prior to contactingthe distal ileum of the individual with the ASBTI.

In some embodiments, contacting the distal ileum of an individual inneed thereof with an ASBTI maintains reduced blood and/or plasma glucoselevels in the individual for at least 24 hours compared to blood and/orplasma glucose levels in the individual prior to contacting the distalileum of the individual with the ASBTI.

In some embodiments, the ASBTI and/or the enterendocrine peptideenhancing agent and/or the FXR agonist is administered orally. In someembodiments, the ASBTI and/or the enterendocrine peptide enhancing agentand/or the FXR agonist is administered as an ileal-pH sensitive releaseformulation that delivers the ASBTI and/or the enterendocrine peptideenhancing agent and/or the FXR agonist to the distal ileum, colon and/orrectum of an individual. In some embodiments, the ASBTI and/or theenterendocrine peptide enhancing agent and/or the FXR agonist isadministered as an enterically coated formulation. In some embodiments,oral delivery of an ASBTI and/or an enterendocrine peptide enhancingagent and/or a FXR agonist provided herein can include formulations, asare well known in the art, to provide prolonged or sustained delivery ofthe drug to the gastrointestinal tract by any number of mechanisms.These include, but are not limited to, pH sensitive release from thedosage form based on the changing pH of the small intestine, slowerosion of a tablet or capsule, retention in the stomach based on thephysical properties of the formulation, bioadhesion of the dosage formto the mucosal lining of the intestinal tract, or enzymatic release ofthe active drug from the dosage form. The intended effect is to extendthe time period over which the active drug molecule is delivered to thesite of action (the ileum) by manipulation of the dosage form. Thus,enteric-coated and enteric-coated controlled release formulations arewithin the scope of the present invention. Suitable enteric coatingsinclude cellulose acetate phthalate, polyvinylacetate phthalate,hydroxypropylmethylcellulose phthalate and anionic polymers ofmethacrylic acid and methacrylic acid methyl ester.

In some embodiments of the methods described above, the ASBTI and/or theenterendocrine peptide enhancing agent and/or the FXR agonist isadministered before ingestion of food. In some embodiments of themethods described above, the ASBTI and/or the enterendocrine peptideenhancing agent and/or the FXR agonist is administered less than about60 minutes before ingestion of food. In some embodiments of the methodsdescribed above, the ASBTI and/or the enterendocrine peptide enhancingagent and/or the FXR agonist is administered less than about 30 minutesbefore ingestion of food. In some embodiments of the methods describedabove, the ASBTI and/or the enterendocrine peptide enhancing agentand/or the FXR agonist is administered after ingestion of food.

Provided herein are methods for prevention and/or treatment ofcongestive heart failure, ventricular dysfunction, toxic hypervolemia,polycystic ovary syndrome, inflammatory bowel disease, impaired bowelintegrity, short bowel syndrome, gastritis, peptic ulcer, or irritablebowel disease comprising contacting the distal ileum of an individual inneed thereof with an ASBTI and/or an enterendocrine peptide enhancingagent and/or a FXR agonist. In some embodiments, the methods furthercomprise administration of a DPP-IV inhibitor, a TGR5 agonist, abiguanide, an incretin mimetic, or GLP-1 or an analog thereof. Providedherein are methods for prevention and/or treatment of radiationenteritis comprising contacting the distal ileum of an individual inneed thereof with an ASBTI and/or an enterendocrine peptide enhancingagent and/or a FXR agonist. In some embodiments, the methods furthercomprise administration of a DPP-IV inhibitor, a TGR5 agonist, abiguanide, an incretin mimetic, or GLP-1 or an analog thereof.

Provided herein are compositions for reducing intraenterocyte bile acidsin an individual in need thereof comprising an Apical Sodium-dependentBile Acid Transporter Inhibitor (ASBTI) and/or an enterendocrine peptideenhancing agent and/or a FXR agonist, and a pharmaceutically acceptablecarrier, wherein the ASBTI is delivered or released non-systemically inthe distal ileum of the individual. Provided herein are compositions forreducing necrosis and/or damage to ileal architecture in an individualin need thereof comprising an Apical Sodium-dependent Bile AcidTransporter Inhibitor (ASBTI) and/or an enterendocrine peptide enhancingagent and/or a FXR agonist, and a pharmaceutically acceptable carrier,wherein the ASBTI and/or the enterendocrine peptide enhancing agentand/or the FXR agonist is delivered or released non-systemically in thedistal ileum of the individual. Provided herein are compositions forreducing caloric intake in an individual in need thereof comprising anApical Sodium-dependent Bile Acid Transporter Inhibitor (ASBTI) and/oran enterendocrine peptide enhancing agent and/or a FXR agonist, and apharmaceutically acceptable carrier, wherein the ASBTI and/or theenterendocrine peptide enhancing agent and/or the FXR agonist isdelivered or released non-systemically in the distal ileum of theindividual. Provided herein are compositions for reducing circulatingblood and/or plasma glucose levels in an individual in need thereofcomprising an Apical Sodium-dependent Bile Acid Transporter Inhibitor(ASBTI) and/or an enterendocrine peptide enhancing agent and/or a FXRagonist, and a pharmaceutically acceptable carrier, wherein the ASBTI isdelivered or released non-systemically in the distal ileum of theindividual. Provided herein are compositions for increasing insulinsecretion in an individual in need thereof comprising an ApicalSodium-dependent Bile Acid Transporter Inhibitor (ASBTI) and/or anenterendocrine peptide enhancing agent and/or a FXR agonist, and apharmaceutically acceptable carrier, wherein the ASBTI is delivered orreleased non-systemically in the distal ileum of the individual.Provided herein are compositions for preventing pancreatic and othercancers comprising contacting the distal ileum of an individual in needthereof with an ASBTI. In any of the aforementioned embodiments, thecompositions further comprise a DPP-IV inhibitor, a TGR5 agonist, abiguanide, an incretin mimetic, or GLP-1 or an analog thereof.

Provided herein, in some embodiments, are ASBTIs for reducingintraenterocyte bile acids, reducing necrosis and/or damage to ilealarchitecture, reducing food intake (caloric intake), or for reducingcirculating blood or plasma glucose levels wherein the ASBTI is notabsorbed systemically following oral administration. In some of suchembodiments, the ASBTI is a compound of Formula I, II, III, IV, V or VIas described herein. In some of such embodiments, the ASBTI is preventedfrom being absorbed in the stomach by its presence in a formulation thatreleases it in the ileum. In some of such embodiments, the ASBTI isadministered in combination with a second therapeutic agent selectedfrom a DPP-IV inhibitor, a biguanide, a thiazolidinedione, an incretinmimetic, GLP-1 or an analog thereof, or a TGR5 agonist.

Provided in some embodiments herein is a kit comprising any compositiondescribed herein (e.g., a pharmaceutical composition formulated forrectal administration) and a device for localized delivery within therectum or colon. In certain embodiments, the device is a syringe, bag,or a pressurized container.

BRIEF DESCRIPTION OF FIGURES

FIG. 1 illustrates the change in plasma glucose level in diabetic db/dbmice after oral administration of a combination of the ASBTI(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideat doses of 0, 0.1, 1, 10 mg/kg and 30 mg/kg sitagliptin (DPP-IVinhibitor) and the ASBTI alone.

FIG. 2 illustrates the change in plasma glucose level in diabetic db/dbmice after oral administration of a combination of the ASBTI1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate at doses of 0, 3, 30, 100 mg/kg and 30 mg/kgsitagliptin (DPP-IV inhibitor).

FIG. 3 illustrates the change in plasma glucose level in diabetic db/dbmice after oral administration of a combination of metformin at doses of0, 3, 30, 100, 300 mg/kg and 30 mg/kg sitagliptin (DPP-IV inhibitor).

FIG. 4 illustrates the change in plasma glucose level in diabetic db/dbmice after oral administration of the ASBTI1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethanesulfonate salt at doses of 0, 3, 30, and 100 mg/kg.

FIG. 5 illustrates a time course for dose-dependent increase in plasmaGLP-1 level in normal rats upon administration of a combination ofmetformin (0, 30, 100, 300 mg/kg) and 30 mg/kg sitagliptin (DPP-IVinhibitor).

FIG. 6 illustrates a dose-dependent increase in plasma GLP-1 level innormal rats upon administration of a combination of the ASBTI1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate (100B) (0, 3, 30, 100 mg/kg) and 30 mg/kg sitagliptin(DPP-IV inhibitor) and the ASBTI alone.

FIG. 7A and FIG. 7B illustrate a time course for dose-dependent increasein plasma GLP-1 level in normal rats upon administration of acombination of the ASBTI1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate (0, 3, 30, 100 mg/kg) and 30 mg/kg sitagliptin (DPP-IVinhibitor) and the ASBTI alone.

FIG. 8 illustrates the change in body weight 24 h after administrationof ASBTI1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate (100B) at doses of 0, 3, 30, 100 mg/kg in combinationwith 30 mg/kg sitagliptin (DPP-IV inhibitor).

FIG. 9 illustrates the change in body weight 24 h after administrationof the ASBTI(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxide(100A) at doses of 0, 0.1, 1, 10 mg/kg in combination with 30 mg/kgsitagliptin (DPP-IV inhibitor) and the ASBTI alone.

FIG. 10 illustrates (1) a comparison of orally administered combinationof(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxide(100A) at doses of 0, 0.1, 1, 10 mg/kg and 30 mg/kg sitagliptin (DPP-IVinhibitor) on blood glucose levels in diabetic db/db mice at 3 hoursafter dose; and (2) a comparison of orally administered combination of1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethanesulfonate salt (100B) at doses of 0, 3, 30, 100 mg/kg and 30mg/kg sitagliptin (DPP-IV inhibitor) on blood glucose levels in diabeticdb/db mice at 3 hours after dose; versus (3) subcutaneously injectedexenatide at doses of 0.02, 0.2 and 2 mg·kg on blood glucose levels indiabetic db/db mice at 3 hours after dose.

FIG. 11A and FIG. 11B illustrate the response of enteroendocrinepeptides to administration of bile salts. FIG. 11A illustrates theplasma PYY concentrations in the caecum, transverse colon, and sigmoidas a result of bile salt administration. FIG. 11B illustrates the plasmaenteroglucagon concentrations in the caecum, transverse colon, andsigmoid as a result of bile salt administration.

FIG. 12 illustrates the affect on food intake of pump infusion of TCA.

FIG. 13 illustrates the affect on body weight of pump infusion of TCA.

FIG. 14 illustrates the affect of the viscosity of a formulationdescribed herein on the food intake of a subject.

FIG. 15 illustrates the cumulative food intake of rats with (dottedlines) and without (solid lines) rectal administration of taurocholate.

FIG. 16 illustrates the cumulative food intake of rats with rectaladministration of three concentrations of taurocholate and thecumulative food intake of rats without rectal administration oftaurocholate.

FIG. 17 illustrates the dose response in rats for the anorectic effectof rectally administered taurocholate.

FIG. 18 illustrates the normal GI distribution of PYY (pmol/g).

FIG. 19 illustrates the acute release of GLP-1 in response to rectaladministration of bile acids in obese diabetic humans treated with aDPP4 inhibitor.

FIG. 20 illustrates the acute release of PYY in response to rectaladministration of bile acids in obese diabetic humans treated with aDPP4 inhibitor.

FIG. 21 illustrates the reduction of spontaneous food intake in responseto rectal administration of bile acids in obese diabetic humans treatedwith a DPP4 inhibitor.

FIG. 22 illustrates the acute release of insulin and in response torectal administration of bile acids in obese diabetic humans treatedwith a DPP4 inhibitor.

FIG. 23 illustrates the incremental integrated response of insulin torectal administration of bile acids in obese diabetic humans treatedwith a DPP4 inhibitor.

FIG. 24 illustrates the acute reduction of plasma glucose concentrationin response to rectal administration of bile acids in obese diabetichumans treated with a DPP4 inhibitor.

FIG. 25 illustrates the incremental integrated response of plasmaglucose concentration to rectally administered bile acids in obesediabetic humans treated with a DPP4 inhibitor.

FIG. 26 illustrates delivery of bile acid salts to the lower gut via abiliary shunt.

DETAILED DESCRIPTION OF THE INVENTION

Described herein is the use of inhibitors of the Apical Sodium-dependentBile Transporter (ASBT) or any recuperative bile salt transporter thatare active in the gastrointestinal (GI) tract.

In certain embodiments, the methods provided herein are utilized inmethods for treating or preventing an inflammatory intestinal conditionor induction of weight loss in an individual in need thereof. In certainembodiments, the methods provided herein comprise administering atherapeutically effective amount of an ASBT inhibitor (ASBTI) and/or anenteroendocrine peptide enhancing agent and/or a FXR agonist to anindividual in need thereof. In some embodiments, such ASBT inhibitorsand/or enteroendocrine peptide enhancing agents and/or FXR agonists arenot systemically absorbed. In some embodiments, such ASBT inhibitorsand/or enteroendocrine peptide enhancing agents and/or FXR agonists aresystemically absorbed. In some of such embodiments, such bile salttransport inhibitors include a moiety or group that prevents, reduces orinhibits the systemic absorption of the compound in vivo. In someembodiments, a charged moiety or group on the compounds prevents,reduces or inhibits the compounds from leaving the gastrointestinaltract and reduces the risk of side effects due to systemic absorption.In some embodiments, the ASBTIs are formulated for delivery to thedistal ileum. In some embodiments, an ASBTI and/or an enteroendocrinepeptide enhancing agent and/or a FXR agonist is minimally absorbed. Insome embodiments, an ASBTI and/or an enteroendocrine peptide enhancingagent and/or a FXR agonist is non-systemically administered to the colonor the rectum of an individual in need thereof. In some embodiments,less than 50%, less than 40%, less than 30%, less than 20%, less than10%, less than 9%, less than 8%, less than 7%, less than 6%, less than5%, less than 4%, less than 3%, less than 2%, or less than 1% of theASBTI and/or the enteroendocrine peptide enhancing agent and/or a FXRagonist is systemically absorbed. In certain embodiments, ASBTIsdescribed herein inhibit scavenging of bile salts by recuperative bileacid salt transporters in the distal gastrointestinal tract (e.g., thedistal ileum, the colon and/or the rectum).

In some instances, the inhibition of bile salt recycling results inhigher concentrations of bile salts in the lumen of the distalgastrointestinal tract or portions thereof (e.g., the distal small boweland/or colon and/or rectum). As used herein, the distal gastrointestinaltract includes the region from the distal ileum to the anus. In someembodiments, the compounds described herein reduce intraenterocyte bileacids or accumulation thereof. In some embodiments, the compoundsdescribed herein reduce necrosis and/or damage to ileal architecture. Incertain embodiments, the higher concentration of bile salts in thedistal small bowel and/or colon and/or rectum modulates (e.g., enhances)the secretion of enteroendocrine peptides in the distal gastrointestinaltract. In some embodiments, the compounds described herein enhance thesecretion of enteroendocrine peptides (e.g., GLP-1, GLP-2,oxyntomodulin, PYY, or a combination thereof) from L-cells that arepresent in the distal ileum, colon and/or the rectum. In certainembodiments, the enhanced secretion of L-cell enteroendocrine peptidesmodulates (e.g., slows or inhibits) gastric emptying and gastric acidsecretion. In certain instances the enhanced secretion of L-cellenteroendocrine peptides induces a feeling of satiety. In someembodiments, the enhanced secretion of L-cell enteroendocrine peptidesreduces food intake thereby inducing weight loss.

Also described herein is the use of such compounds for reducing ormaintaining weight in individuals (e.g., individuals interested inlosing weight, reducing weight, and/or maintaining a healthy body weightand lifestyle). In certain instances, the compounds described herein areuseful in the treatment of diseases or conditions that are modulated byenteroendocrine peptides secreted by L-cells of the distalgastrointestinal tract, including L-cells in the distal ileum, the colonand/or the rectum. In some instances, enhanced enteroendocrine peptidesecretion (e.g., increase in GLP-1 secretion) reduces blood or plasmaglucose levels. In some embodiments, the compounds described herein arealso useful in the treatment of metabolic disorders (e.g., diabetes,metabolic syndrome or the like) because they do not cause side effects(e.g., weight gain) that are associated with conventional therapies formetabolic disorders.

Compounds

In some embodiments, provided herein are ASBT inhibitors that reduce orinhibit bile acid recycling in the distal gastrointestinal (GI) tract,including the distal ileum, the colon and/or the rectum. In certainembodiments, the ASBTIs are systemically absorbed. In certainembodiments, the ASBTIs are not systemically absorbed. In someembodiments, ASBTIs described herein are modified or substituted (e.g.,with a -L-K group) to be non-systemic. In certain embodiments, any ASBTinhibitor is modified or substituted with one or more charged groups(e.g., K) and optionally, one or more linker (e.g., L), wherein L and Kare as defined herein.

In some embodiments, an ASBTI suitable for the methods described hereinis a compound of Formula I:

wherein:R¹ is a straight chained C₁₋₆ alkyl group;R² is a straight chained C₁₋₆ alkyl group;R³ is hydrogen or a group OR¹¹ in which R¹¹ is hydrogen, optionallysubstituted C₁₋₆ alkyl or a C₁₋₆ alkylcarbonyl group;R⁴ is pyridyl or optionally substituted phenyl or -L_(z)-K_(z); whereinz is 1, 2 or 3; each L is independently a substituted or unsubstitutedalkyl, a substituted or unsubstituted heteroalkyl, a substituted orunsubstituted alkoxy, a substituted or unsubstituted aminoalkyl group, asubstituted or unsubstituted aryl, a substituted or unsubstitutedheteroaryl, a substituted or unsubstituted cycloalkyl, or a substitutedor unsubstituted heterocycloalkyl; each K is a moiety that preventssystemic absorption;R⁵, R⁶, R⁷ and R⁸ are the same or different and each is selected fromhydrogen, halogen, cyano, R⁵-acetylide, OR¹⁵, optionally substitutedC₁₋₆ alkyl, COR¹⁵, CH(OH)R¹⁵, S(O)_(n)R¹⁵, P(O)(OR¹⁵)₂, OCOR¹⁵, OCF3,OCN, SCN, NHCN, CH₂OR¹⁵, CHO, (CH₂)_(p)CN, CONR¹²R¹³, (CH₂)_(p)CO₂R¹⁵,(CH₂)_(p)NR¹²R¹³, CO₂R¹⁵, NHCOCF₃, NHSO₂R¹⁵, OCH₂OR¹⁵, OCH═CHR¹⁵,O(CH₂CH₂O)R¹⁵, O(CH₂)_(p)SO₃R¹⁵, O(CH₂)_(p)NR¹²R¹³,O(CH₂)_(p)N⁺R¹²R¹³R¹⁴ and —W—R³¹, wherein W is O or NH and R³¹ isselected from

wherein p is an integer from 1-4, n is an integer from 0-3 and, R¹²,R¹³, R¹⁴ and R¹⁵ are independently selected from hydrogen and optionallysubstituted C₁₋₆ alkyl; orR⁶ and R⁷ are linked to form a group

wherein R¹² and R¹³ are as hereinbefore defined and m is 1 or 2; and

R⁹ and R¹⁰ are the same or different and each is selected from hydrogenor C₁₋₆ alkyl; and salts, solvates and physiologically functionalderivatives thereof.

In some embodiments of the methods, the compound of Formula I is acompound wherein

R¹ is a straight chained C₁₋₆ alkyl group;

R² is a straight chained C₁₋₆ alkyl group;

R³ is hydrogen or a group OR¹¹ in which R¹¹ is hydrogen, optionallysubstituted C₁₋₆ alkyl or a C₁₋₆ alkylcarbonyl group;

R⁴ is optionally substituted phenyl;

R⁵, R⁶ and R⁸ are independently selected from hydrogen, C₁₋₄ alkyloptionally substituted by fluorine, C₁₋₄ alkoxy, halogen, or hydroxy;

R⁷ is selected from halogen, cyano, R¹⁵-acetylide, OR¹⁵, optionallysubstituted C₁₋₆ alkyl, COR¹⁵, CH(OH)R¹⁵, S(O)_(n)R¹⁵, P(O)(OR¹⁵)₂,OCOR¹⁵, OCF₃, OCN, SCN, HNCN, CH₂OR¹⁵, CHO, (CH₂)_(p)CN, CONR¹²R¹³,(CH₂)_(p)CO₂R¹⁵, (CH₂)_(p)NR¹²R¹³, CO₂R¹⁵, NHCOCF₃, NHSO₂R¹⁵, OCH₂OR¹⁵,OCH═CHR¹⁵, O(CH₂CH₂O)R¹⁵, O(CH₂)_(p)SO₃R¹⁵, O(CH₂)_(p)NR¹²R¹³ andO(CH₂)_(p)N⁺R¹²R¹³R¹⁴;

wherein n, p and R¹² to R¹⁵ are as hereinbefore defined;

with the proviso that at least two of R⁵ to R⁸ are not hydrogen; and

salts solvates and physiologically functional derivatives thereof.

In some embodiments of the methods described herein, the compound ofFormula I is a compound wherein

R¹ is a straight chained C₁₋₆ alkyl group;

R² is a straight chained C₁₋₆ alkyl group;

R³ is hydrogen or a group OR¹¹ in which R¹¹ is hydrogen, optionallysubstituted C₁₋₆ alkyl or a C₁₋₆ alkylcarbonyl group;

R⁴ is un-substituted phenyl;

R⁵ is hydrogen or halogen;

R⁶ and R⁸ are independently selected from hydrogen, C₁₋₄ alkyloptionally substituted by fluorine, C₁₋₄ alkoxy, halogen, or hydroxy;

R⁷ is selected from OR¹⁵, S(O)_(n)R¹⁵, OCOR¹⁵, OCF₃, OCN, SCN, CHO,OCH₂OR¹⁵, OCH═CHR¹⁵, O(CH₂CH₂O)_(n)R¹⁵, O(CH₂)_(p)SO₃R¹⁵,O(CH₂)_(p)NR¹²R¹³ and O(CH₂)_(p)N⁺R¹²R¹³R¹⁴ wherein p is an integer from1-4, n is an integer from 0-3, and R¹², R¹³, R¹⁴, and R¹⁵ areindependently selected from hydrogen and optionally substituted C₁₋₆alkyl;R⁹ and R¹⁰ are the same or different and each is selected from hydrogenor C₁₋₆ alkyl; and salts, solvates and physiologically functionalderivatives thereof.

In some embodiments of the methods, wherein the compound of Formula I isa compound wherein

R¹ is methyl, ethyl or n-propyl;

R² is methyl, ethyl, n-propyl, n-butyl or n-pentyl;

R³ is hydrogen or a group OR¹¹ in which R¹¹ is hydrogen, optionallysubstituted C₁₋₆ alkyl or a C₁₋₆ alkylcarbonyl group;

R⁴ is un-substituted phenyl;

R⁵ is hydrogen;

R⁶ and R⁸ are independently selected from hydrogen, C₁₋₄ alkyloptionally substituted by fluorine, C₁₋₄ alkoxy, halogen, or hydroxy;

R⁷ is selected from OR¹⁵, S(O)_(n)R¹⁵, OCOR¹⁵, OCF₃, OCN, SCN, CHO,OCH₂OR¹⁵, OCH═CHR¹⁵, O(CH₂CH₂O)_(n)R¹⁵, O(CH₂)_(p)SO₃R¹⁵,O(CH₂)_(p)NR¹²R¹³ and O(CH₂)_(p)N⁺R¹²R¹³R¹⁴ wherein p is an integer from1-4, n is an integer from 0-3, and R¹², R¹³, R¹⁴, and R¹⁵ areindependently selected from hydrogen and optionally substituted C₁₋₆alkyl;R⁹ and R¹⁰ are the same or different and each is selected from hydrogenor C₁₋₆ alkyl; and salts, solvates and physiologically functionalderivatives thereof.

In some embodiments of the methods, the compound of Formula I is acompound wherein

R¹ is methyl, ethyl or n-propyl;

R² is methyl, ethyl, n-propyl, n-butyl or n-pentyl;

R³ is hydrogen or a group OR¹¹ in which R¹¹ is hydrogen, optionallysubstituted C₁₋₆ alkyl or a C₁₋₆ alkylcarbonyl group;

R⁴ is un-substituted phenyl;

R⁵ is hydrogen;

R⁶ is C₁₋₄ alkoxy, halogen, or hydroxy;

R⁷ is OR¹⁵, wherein R¹⁵ is hydrogen or optionally substituted C₁₋₆alkyl;

R⁸ is hydrogen or halogen;

R⁹ and R¹⁰ are the same or different and each is selected from hydrogenor C₁₋₆ alkyl; and salts, solvates and physiologically functionalderivatives thereof.

In some embodiments of the methods, the compound of Formula I is

-   (3R,5R)-3-Butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine    1,1-dioxide;-   (3R,5R)-3-Butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepin-4-ol    1,1-dioxide;-   (±)-Trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine    1,1-dioxide;-   (±)-Trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepin-4-ol    1,1-dioxide;-   (3R,5R)-7-Bromo-3-butyl-3-ethyl-2,3,4,5-tetrahydro-8-methoxy-5-phenyl-1,4-benzothiazepine    1,1-dioxide;-   (3R,5R)-7-Bromo-3-butyl-3-ethyl-2,3,4,5-tetrahydro-8-methoxy-5-phenyl-1,4-benxothiaxepin-4-ol    1,1-dioxide;-   (3R,5R)-3-Butyl-3-ethyl-2,3,4,5-tetrahydro-5-phenyl-1,4-benzothiazepine-7,8-diol    1,1-dioxide;-   (3R,5R)-3-Butyl-3-ethyl-2,3,4,5-tetrahydro-8-methoxy-5-phenyl-1,4-benzothiazepin-7-ol    1,1-dioxide;-   (3R,5R)-3-Butyl-3-ethyl-2,3,4,5-tetrahydro-7-methoxy-5-phenyl-1,4-benzothiazepin-8-ol    1,1-dioxide;-   (±)-Trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-8-methoxy-5-phenyl-1,4-benzothiazepine    1,1-dioxide;-   (±)-Trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-5-phenyl-1,4-benzothiazepin-8-ol    1,1-dioxide;-   (±)-Trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-5-phenyl-1,4-benzothiazepine-4,8-diol;-   (±)-Trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-5-phenyl-1,4-benzothiazepin-8-thiol    1,1-dioxide;-   (±)-Trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-5-phenyl-1,4-benzothiazepin-8-sulfonic    acid 1,1-dioxide;-   (±)-Trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-8,9-dimethoxy-5-phenyl-1,4-benzothiazepine    1,1-dioxide;-   (3R,5R)-3-butyl-7,8-diethoxy-2,3,4,5-tetrahydro-5-phenyl-1,4-benzothiazepine    1,1-dioxide;-   (±)-Trans-3-butyl-8-ethoxy-3-ethyl-2,3,4,5-tetrahydro-5-phenyl-1,4-benzothiazepine    1,1-dioxide;-   (+)-Trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-8-isopropoxy-5-phenyl-1,4-benzothiazepine    1,1-dioxide hydrochloride;-   (±)-Trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-5-phenyl-1,4-benzothiazepin-8-carbaldehyde-1,1-dioxide;-   3,3-Diethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine    1,1-dioxide;-   3,3-Diethyl-2,3,4,5-tetrahydro-8-methoxy-5-phenyl-1,4-benzothiazepine    1,1-dioxide;-   3,3-Diethyl-2,3,4,5-tetrahydro-5-phenyl-1,4-benzothiazpin-4,8-diol    1,1-dioxide;-   (RS)-3,3-Diethyl-2,3,4,5-tetrahydro-4-hydroxy-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine    1,1-dioxide;-   (+)-Trans-3-butyl-8-ethoxy-3-ethyl-2,3,4,5-tetrahydro-5-phenyl-1,4-benzothiazepin-4-ol-1-dioxide;-   (±)-Trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-8-isopropoxy-5-phenyl-1,4-benzothiazepin-4-ol    1,1-dioxide;-   (+)-Trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8,9-trimethoxy-5-phenyl-1,4-benzothiazepin-4-ol    1,1-dioxide;-   (3R,5R)-3-butyl-3-ethyl-2,3,4,5-tetrahydro-5-phenyl-1,4-benzothiazepin-4,7,8-triol    1,1-dioxide;-   (±)-Trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-4,7,8-trimethoxy-5-phenyl-1,4-benzothiazepine    1,1-dioxide;-   3,3-Diethyl-2,3,4,5-tetrahydro-5-phenyl-1,4-benzothiazepin-8-ol    1,1-dioxide;-   3,3-Diethyl-2,3,4,5-tetrahydro-7-methoxy-5-phenyl-1,4-benzothiazepin-8-ol    1,1-dioxide;-   3,3Dibutyl-2,3,4,5-tetrahydro-5-phenyl-1,4-benzothiazepin-8-ol    1,1-dioxide;-   (+)-Trans-3-Butyl-3-ethyl-2,3,4,5-tetrahydro-1,1-dioxo-5-phenyl-1,4-benzothiazepin-8-yl    hydrogen sulfate;-   or-   3,3-Diethyl-2,3,4,5-tetrahydro-1,1-dioxo-5-phenyl-1,4-benzothiazepin-8-yl    hydrogen sulfate.

In some embodiments, the compound of Formula I is

In some embodiments of the methods, the compound of Formula I is

In some embodiments, an ASBTI suitable for the methods described hereinis a compound of Formula II

wherein:

-   -   q is an integer from 1 to 4;    -   n is an integer from 0 to 2;    -   R¹ and R² are independently selected from the group consisting        of H, alkyl, alkenyl, alkynyl, haloalkyl, alkylaryl, arylalkyl,        alkoxy, alkoxyalkyl, dialkylamino, alkylthio, (polyalkyl)aryl,        and cycloalkyl,    -   wherein alkyl, alkenyl, alkynyl, haloalkyl, alkylaryl,        arylalkyl, alkoxy, alkoxyalkyl, dialkylamino, alkylthio,        (polyalkyl)aryl, and cycloalkyl optionally are substituted with        one or more substituents selected from the group consisting of        OR⁹, NR⁹R¹⁰, N⁺R⁹R¹⁰R^(w)A⁻, S⁺R⁹, S⁺R⁹R¹⁰A⁻, P⁺R⁹R¹⁰R¹¹A⁻,        S(O)R⁹, SO₂R⁹, SO₃R⁹, CO₂R⁹, CN, halogen, oxo, and CONR⁹R¹⁰,    -   wherein alkyl, alkenyl, alkynyl, alkylaryl, alkoxy, alkoxyalkyl,        (polyalkyl)aryl, and cycloalkyl optionally have one or more        carbons replaced by O, NR⁹, N⁺R⁹R¹⁰A⁻, S, SO, SO₂, S⁺R⁹A⁻,        P⁺R⁹R¹⁰A⁻, or phenylene,    -   wherein R⁹, R¹⁰, and R^(w) are independently selected from the        group consisting of H, alkyl, alkenyl, alkynyl, cycloalkyl,        aryl, acyl, heterocycle, ammoniumalkyl, arylalkyl, and        alkylammoniumalkyl; or    -   R¹ and R² taken together with the carbon to which they are        attached form C₃-C₁₀ cycloalkyl;    -   R³ and R⁴ are independently selected from the group consisting        of H, alkyl, alkenyl, alkynyl, acyloxy, aryl, heterocycle, OR⁹,        NR⁹R¹⁰, SR⁹, S(O)R⁹, SO₂R⁹, and SO₃R⁹, wherein R⁹ and R¹⁰ are as        defined above; or    -   R³ and R⁴ together ═O, ═NOR¹¹, ═S, ═NNR¹¹R¹², ═NR⁹, or ═CR¹¹R¹²,    -   wherein R¹¹ and R¹² are independently selected from the group        consisting of H, alkyl, alkenyl, alkynyl, aryl, arylalkyl,        alkenylalkyl, alkynylalkyl, heterocycle, carboxyalkyl,        carboalkoxyalkyl, cycloalkyl, cyanoalkyl, OR⁹, NR⁹R¹⁰, SR⁹,        S(O)R⁹, SO₂R⁹, SO₃R⁹, CO₂R⁹, CN, halogen, oxo, and CONR⁹R¹⁰,        wherein R⁹ and R¹⁰ are as defined above, provided that both R³        and R⁴ cannot be OH, NH₂, and SH, or    -   R¹¹ and R¹² together with the nitrogen or carbon atom to which        they are attached form a cyclic ring;        -   R⁵ and R⁶ are independently selected from the group            consisting of H, alkyl, alkenyl, alkynyl, aryl, cycloalkyl,            heterocycle, quaternary heterocycle, quarternary heteroaryl,            OR³⁰, SR⁹, S(O)R⁹, SO₂R⁹, SO₃R⁹, and -L_(z)-K_(z);        -   wherein z is 1, 2 or 3; each L is independently a            substituted or unsubstituted alkyl, a substituted or            unsubstituted heteroalkyl, a substituted or unsubstituted            alkoxy, a substituted or unsubstituted aminoalkyl group, a            substituted or unsubstituted aryl, a substituted or            unsubstituted heteroaryl, a substituted or unsubstituted            cycloalkyl, or a substituted or unsubstituted            heterocycloalkyl; each K is a moiety that prevents systemic            absorption;        -   wherein alkyl, alkenyl, alkynyl, aryl, cycloalkyl,            heterocycle, quaternary heterocycle, and quaternary            heteroaryl can be substituted with one or more substituent            groups independently selected from the group consisting of            alkyl, alkenyl, alkynyl, polyalkyl, polyether, aryl,            haloalkyl, cycloalkyl, heterocycle, arylalkyl, quaternary            heterocycle, quaternary heteroaryl, halogen, oxo, OR¹³,            NR¹³R¹⁴, SR¹³, S(O)R¹³, SO₂R¹³, SO₃R¹³, NR¹³OR¹⁴,            NR¹³NR¹⁴R¹⁵, NO₂, CO₂R¹³, CN, OM, SO₂OM, SO₂NR¹³R¹⁴,            C(O)NR¹³R¹⁴, C(O)OM, CR¹³, P(O)R¹³R¹⁴, P⁺R¹³R¹⁴R¹⁵A⁻,            P(OR¹³)OR¹⁴, S⁺R¹³R¹⁴A⁻, and N⁺R⁹R¹¹R¹²A⁻,            wherein:    -   A⁻ is a pharmaceutically acceptable anion and M is a        pharmaceutically acceptable cation, said alkyl, alkenyl,        alkynyl, polyalkyl, polyether, aryl, haloalkyl, cycloalkyl, and        heterocycle can be further substituted with one or more        substituent groups selected from the group consisting of OR⁷,        NR⁷R⁸, S(O)R⁷, SO₂R⁷, SO₃R⁷, CO₂R⁷, CN, oxo, CONR⁷R⁸,        N⁺R⁷R⁸R⁹A⁻, alkyl, alkenyl, alkynyl, aryl, cycloalkyl,        heterocycle, arylalkyl, quaternary heterocycle, quaternary        heteroaryl, P(O)R⁷R⁸, P⁺R⁷R⁸R⁹A⁻, and P(O)(OR⁷) OR⁸ and    -   wherein said alkyl, alkenyl, alkynyl, polyalkyl, polyether,        aryl, haloalkyl, cycloalkyl, and heterocycle can optionally have        one or more carbons replaced by O, NR⁷, N⁺R⁷R⁸A⁻, S, SO, SO₂,        S⁺R⁷A⁻, PR⁷, P(O)R⁷, P⁺R⁷R⁸A⁻, or phenylene, and R¹³, R¹⁴, and        R¹⁵ are independently selected from the group consisting of        hydrogen, alkyl, alkenyl, alkynyl, polyalkyl, aryl, arylalkyl,        cycloalkyl, heterocycle, heteroaryl, quaternary heterocycle,        quaternary heteroaryl, and quaternary heteroarylalkyl,    -   wherein alkyl, alkenyl, alkynyl, arylalkyl, heterocycle, and        polyalkyl optionally have one or more carbons replaced by O,        NR⁹, N⁺R⁹R¹⁰A⁻, S, SO, SO₂, S⁺R⁹A⁻, PR, P⁺R⁹R¹⁰A⁻, P(O)R⁹,        phenylene, carbohydrate, amino acid, peptide, or polypeptide,        and    -   R¹³, R¹⁴ and R¹⁵ are optionally substituted with one or more        groups selected from the group consisting of sulfoalkyl,        quaternary heterocycle, quaternary heteroaryl, OR⁹, NR⁹R¹⁰,        N⁺R⁹R¹⁰R¹¹R¹²A⁻, SR⁹, S(O) R⁹, SO₂R⁹, SO₃R⁹, oxo, CO₂R⁹, CN,        halogen, CONR⁹R¹⁰, SO₂OM, SO₂NR⁹R¹⁰, PO(OR¹⁶)OR¹⁷, P⁺R⁹R⁹R¹⁰A⁻,        S⁺R⁹R¹⁰A⁻, and C(O)OM,    -   wherein R¹⁶ and R¹⁷ are independently selected from the        substituents constituting R⁹ and M; or    -   R¹⁴ and R¹⁵, together with the nitrogen atom to which they are        attached, form a cyclic ring; and    -   is selected from the group consisting of alkyl, alkenyl,        alkynyl, cycloalkyl, aryl, acyl, heterocycle, ammoniumalkyl,        alkylammoniumalkyl, and arylalkyl; and    -   R⁷ and R⁸ are independently selected from the group consisting        of hydrogen and alkyl; and    -   one or more R^(x) are independently selected from the group        consisting of H, alkyl, alkenyl, alkynyl, polyalkyl, acyloxy,        aryl, arylalkyl, halogen, haloalkyl, cycloalkyl, heterocycle,        heteroaryl, polyether, quaternary heterocycle, quaternary        heteroaryl, OR¹³, NR¹³R¹⁴, SR¹³, S(O)R¹³, S(O)₂R¹³, SO₃R¹³,        S⁺R¹³R¹⁴A⁻, NR¹³OR¹⁴, NR¹³NR¹⁴R¹⁵, NO₂, CO₂R¹³, CN, OM, SO₂OM,        SO₂NR¹³R¹⁴, NR¹⁴C(O)R¹³, C(O)NR¹³R¹⁴, NR¹⁴C(O)R¹³, C(O)OM,        COR¹³, OR¹⁸, S(O)_(n)NR¹⁸, NR¹³R¹⁸, NR¹⁸R¹⁴, N⁺12⁹R¹¹R¹²A⁻,        P⁺R⁹R¹⁰R¹¹R¹²A⁻, amino acid, peptide, polypeptide, and        carbohydrate,    -   wherein alkyl, alkenyl, alkynyl, cycloalkyl, aryl, polyalkyl,        heterocycle, acyloxy, arylalkyl, haloalkyl, polyether,        quaternary heterocycle, and quaternary heteroaryl can be further        substituted with OR⁹, NR⁹R¹⁰, N⁺R⁹R¹¹R¹²A⁻, SR⁹, S(O)R⁹, SO₂R⁹,        SO₃R⁹, oxo, CO₂R⁹, CN, halogen, CONR⁹R⁰, SO₂OM, SO₂NR⁹R¹⁰,        PO(OR¹⁶)OR¹⁷, P⁺R⁹R¹⁰R¹¹R¹²A⁻, S⁺R⁹R¹⁰A⁻, or C(O)M, and    -   wherein R¹⁸ is selected from the group consisting of acyl,        arylalkoxycarbonyl, arylalkyl, heterocycle, heteroaryl, alkyl,    -   wherein acyl, arylalkoxycarbonyl, arylalkyl, heterocycle,        heteroaryl, alkyl, quaternary heterocycle, and quaternary        heteroaryl optionally are substituted with one or more        substituents selected from the group consisting of OR⁹, NR⁹R¹⁰,        N⁺R⁹R¹¹R¹²A⁻, SR⁹, S(O)R⁹, SO₂R⁹, SO₃R⁹, OXO, CO₃R⁹, CN,        halogen, CONR⁹R¹⁰, SO₃R⁹, SO₂OM, SO₂NR⁹R¹⁰, PO(OR¹⁶)OR¹⁷, and        C(O)OM,    -   wherein in R^(x), one or more carbons are optionally replaced by        O, NR¹³, N⁺R¹³R¹⁴A⁻, S, SO, SO₂, S⁺R¹³A⁻, PR¹³, P(O)R¹³,        P⁺R¹³R¹⁴A⁻, phenylene, amino acid, peptide, polypeptide,        carbohydrate, polyether, or polyalkyl,    -   wherein in said polyalkyl, phenylene, amino acid, peptide,        polypeptide, and carbohydrate, one or more carbons are        optionally replaced by O, NR⁹, R⁹R¹⁰A⁻, S, SO, SO₂, S⁺R⁹A⁻, PR⁹,        P⁺R⁹R¹⁰A⁻, or P(O)R⁹;    -   wherein quaternary heterocycle and quaternary heteroaryl are        optionally substituted with one or more groups selected from the        group consisting of alkyl, alkenyl, alkynyl, polyalkyl,        polyether, aryl, haloalkyl, cycloalkyl, heterocycle, arylalkyl,        halogen, oxo, OR¹³, NR¹³R¹⁴, SR¹³, S(O)R¹³, SO₂R, SO₃R¹³,        NR¹³OR¹⁴, NR¹³NR¹⁴R¹⁵, NO₂, CO₂R¹³, CN, OM, SO₂OM, SO₂NR¹³R¹⁴,        C(O)NR¹³R¹⁴, C(O)OM, COR¹³, P(O)R¹³R¹⁴, P⁺R¹³R¹⁴R¹⁵A⁻,        P(OR¹³)OR¹⁴, S⁺R¹³R¹⁴A⁻, and N⁺R⁹R¹¹R¹²A⁻,    -   provided that both R⁵ and R⁶ cannot be hydrogen or SH;    -   provided that when R⁵ or R⁶ is phenyl, only one of R¹ or R² is        H;        provided that when q=1 and R^(x) is styryl, anilido, or        anilinocarbonyl, only one of R⁵ or R⁶ is alkyl; or a        pharmaceutically acceptable salt, solvate, or prodrug thereof.

In some embodiments of the methods, the compound of Formula II is acompound

wherein

R⁵ and R⁶ are independently selected from the group consisting of H,aryl, heterocycle, quaternary heterocycle, and quarternary heteroaryl

-   -   wherein the aryl, heteroaryl, quaternary heterocycle and        quaternary heteroaryl are optionally substituted with one or        more groups selected from the group consisting of alkyl,        alkenyl, alkynyl, polyalkyl, polyether, aryl, haloalkyl,        cycloalkyl, heterocycle, arylalkyl, halogen, oxo, OR¹³, NR¹³R¹⁴,        SR¹³, S(O)R¹³, SO₂R¹³, SO₃R¹³, NR¹³OR¹⁴, NR¹³NR¹⁴R¹⁵, NO₂,        CO₂R¹³, CN, OM, SO₂OM, SO₂NR¹³R¹⁴, C(O)NR¹³R¹⁴, C(O)OM, COR¹³,        P(O)R¹³R¹⁴, P⁺R¹³R¹⁴R¹⁵A⁻, P(OR¹³)OR⁴, S⁺R¹³R¹⁴A⁻, N⁺R⁹R¹¹R¹²A⁻        and -L_(z)-K_(z).

In some embodiments of the methods, the compound of Formula II is acompound

wherein

R⁵ or R⁶ is —Ar—(R^(y))_(t)

-   -   t is an integer from 0 to 5;    -   Ar is selected from the group consisting of phenyl, thiophenyl,        pyridyl, piperazinyl, piperonyl, pyrrolyl, naphthyl, furanyl,        anthracenyl, quinolinyl, isoquinolinyl, quinoxalinyl,        imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, pyrimidinyl,        thiazolyl, triazolyl, isothiazolyl, indolyl, benzoimidazolyl,        benzoxazolyl, benzothiazolyl, and benzoisothiazolyl; and    -   one or more R^(y) are independently selected from the group        consisting of alkyl, alkenyl, alkynyl, polyalkyl, polyether,        aryl, halo alkyl, cycloalkyl, heterocycle, arylalkyl, halogen,        oxo, OR¹³, NR¹³R¹⁴, SR¹³, S(O)R¹³, SO₂R¹³, SO₃R¹³, NR¹³OR⁴,        NR¹³NR⁴R¹⁵, NO₂, CO₂R¹³, CN, OM, SO₂OM, SO₂NR¹³R¹⁴, C(O)NR¹³R¹⁴,        C(O)OM, COR¹³, P(O)R¹³R¹⁴, P⁺R¹³R¹⁴R¹⁵A⁻, P(OR³)OR⁴, S⁺R¹³R¹⁴A⁻,        N⁺R⁹R¹⁰R¹²A⁻ and -L_(z)-K_(z);    -   wherein said alkyl, alkenyl, alkynyl, polyalkyl, polyether,        aryl, haloalkyl, cycloalkyl, and heterocycle can be further        substituted with one or more substituent groups selected from        the group consisting of OR¹³, NR¹³R¹⁴, SR¹³, S(O)R¹³, SO₂R¹³,        SO₃R¹³, NR¹³OR¹⁴, NR¹³NR¹⁴R¹⁵, NO₂, CO₂R¹³, CN, oxo, CONR⁷R⁸,        N⁺R⁷R⁸R⁹A⁻, alkyl, alkenyl, alkynyl, aryl, cycloalkyl,        heterocycle, arylalkyl, quaternary heterocycle, quaternary        heteroaryl, P(O)R⁷R⁸, P⁺R⁷R⁸A⁻, and P(O)(OR⁷)OR⁸, and or        phenylene;    -   wherein said alkyl, alkenyl, alkynyl, polyalkyl, polyether,        aryl, haloalkyl, cycloalkyl, and heterocycle can optionally have        one or more carbons replaced by O, NR⁷, N⁺R⁷R⁸A⁻, S, SO, SO₂,        S⁺R⁷A⁻, PR⁷, P(O)R⁷, P⁺R⁷R⁸A⁻, or phenylene.

In some embodiments of the methods, the compound of Formula II is acompound wherein R⁵ or R⁶ is

In some embodiments of the methods, the compound of Formula II is acompound wherein n is 1 or 2. In some embodiments of the methods, thecompound of Formula II is a compound wherein R¹ and R² are independentlyH or C₁₋₇ alkyl. In some embodiments of the methods, the compound ofFormula II is a compound wherein each C₁₋₇ alkyl is independently ethyl,n-propyl, n-butyl, or isobutyl. In some embodiments of the methods, thecompound of Formula II is a compound wherein R³ and R⁴ are independentlyH or OR⁹. In some embodiments of the methods, compound of Formula II isa compound wherein R⁹ is H

In some embodiments of the methods, the compound of Formula II is acompound wherein one or more R^(x) are in the 7-, 8- or 9-position ofthe benzo ring of Formula II. In some embodiments of the methods, thecompound of Formula II is a compound wherein R^(x) is in the 7-positionof the benzo ring of Formula II. In some embodiments of the methods, thecompound of Formula II is a compound wherein one or more R^(x) areindependently selected from OR¹³ and NR¹³R¹⁴.

In some embodiments of the methods, the compound of Formula II is acompound

wherein:

q is 1 or 2;

n is 2;

R¹ and R² are each alkyl;

R³ is hydroxy;

R⁴ and R⁶ are hydrogen;

R⁵ has the formula

whereint is an integer from 0 to 5;

-   -   one or more R^(Y) are OR¹³;    -   R¹³ is selected from the group consisting of hydrogen, alkyl,        alkenyl, alkynyl, polyalkyl, aryl, arylalkyl, cycloalkyl,        heterocycle, heteroaryl, quaternary heterocycle, quaternary        heteroaryl, and quaternary heteroarylalkyl;    -   said R¹³ alkyl, alkenyl, alkynyl, arylalkyl, heterocycle, and        polyalkyl groups optionally have one or more carbons replaced by        O, NR⁹, N⁺R⁹R¹⁰A⁻, S, SO, SO₂, S⁺R⁹A⁻, PR⁹, P⁺R⁹R¹⁰A⁻, P(O)R⁹,        phenylene, carbohydrate, amino acid, peptide, or polypeptide;    -   R¹³ is optionally substituted with one or more groups selected        from the group consisting of sulfoalkyl, quaternary heterocycle,        quaternary heteroaryl, OR⁹, NR⁹R¹⁰, N⁺R⁹R¹⁰R¹²A⁻, SR⁹, S(O)R⁹,        SO₂R⁹, SO₃R⁹, oxo, CO₂R⁹, CN, halogen, CONR⁹R¹⁰, SO₂OM,        SO₂NR⁹R¹⁰, PO(OR¹⁶)OR⁷, P⁺R⁹R¹⁰R¹¹A⁻, S⁺R⁹R¹⁰A⁻, and C(O)OM,    -   wherein A is a pharmaceutically acceptable anion, and M is a        pharmaceutically acceptable cation,    -   R⁹ and R¹⁰ are independently selected from the group consisting        of H, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, acyl,        heterocycle, ammoniumalkyl, arylalkyl, and alkylammoniumalkyl;    -   R¹¹ and R¹² are independently selected from the group consisting        of H, alkyl, alkenyl, alkynyl, aryl, arylalkyl, alkenylalkyl,        alkynylalkyl, heterocycle, carboxyalkyl, carboalkoxyalkyl,        cycloalkyl, cyanoalkyl, OR⁹, NR⁹R¹⁰, SR⁹, S(O)R⁹, SO₂R⁹, SO₃R⁹,        CO₂R⁹, CN, halogen, oxo, and CONR⁹R¹⁰, wherein R⁹ and R¹⁰ are as        defined above, provided that both R³ and R⁴ cannot be OH, NH₂,        and SH; or    -   R¹¹ and R¹² together with the nitrogen or carbon atom to which        they are attached form a cyclic ring; and    -   R¹⁶ and R¹⁷ are independently selected from the substituents        constituting R⁹ and M;    -   R⁷ and R⁸ are hydrogen; and    -   one or more R^(x) are independently selected from the group        consisting of alkoxy, alkylamino and dialkylamino and —W—R³¹,        wherein W is O or NH and R³¹ is selected from

-   -   or a pharmaceutically acceptable salt, solvate, or prodrug        thereof.

In some embodiments, a compound of Formula II is

or the like.

In some embodiments of the methods, the compound of Formula II is

In certain embodiments, ASBTIs suitable for the methods described hereinare non-systemic analogs of Compound 100C. Certain compounds providedherein are Compound 100C analogues modified or substituted to comprise acharged group. In specific embodiments, the Compound 100C analogues aremodified or substituted with a charged group that is an ammonium group(e.g., a cyclic ar acyclic ammonium group). In certain embodiments, theammonium group is a non-protic ammonium group that contains aquarternary nitrogen.

In some embodiments, a compound of Formula II is

In some embodiments, a compound of Formula II is1-[[5-[[3-[(3S,4R,5R)-3-butyl-7-(dimethylamino)-3-ethyl-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5yl]phenyl]amino]-5-oxopentyl]amino]-1-deoxy-D-glucitolor SA HMR1741 (a.k.a. BARI-1741).

In some embodiments, a compound of Formula II is

In some embodiments, a compound of Formula II is potassium((2R,3R,4S,5R,6R)-4-benzyloxy-6-{3-[3-((3S,4R,5R)-3-butyl-7-dimethylamino-3-ethyl-4-hydroxy-1,1-dioxo-2,3,4,5-tetrahydro-1H-benzo[b]thiepin-5-yl)-phenyl]-ureido}-3,5-dihydroxy-tetrahydro-pyran-2-ylmethyl)sulphateethanolate, hydrate or SAR548304B (a.k.a. SAR-548304).

In some embodiments, an ASBTI suitable for the methods described hereinis a compound of Formula III:

wherein:

-   -   each R¹, R² is independently H, hydroxy, alkyl, alkoxy,        —C(═X)YR⁸, —YC(═X)R⁸, substituted or unsubstituted alkyl,        substituted or unsubstituted heteroalkyl, substituted or        unsubstituted aryl, substituted or unsubstituted alkyl-aryl,        substituted or unsubstituted cycloalkyl, substituted or        unsubstituted alkyl-cycloalkyl, substituted or unsubstituted        heteroaryl, substituted or unsubstituted alkyl-heteroaryl,        substituted or unsubstituted heterocycloalkyl, substituted or        unsubstituted alkyl-heterocycloalkyl, or -L-K; or R¹ and R²        together with the nitrogen to which they are attached form a        3-8-membered ring that is optionally substituted with R⁸;    -   each R³, R⁴ is independently H, hydroxy, alkyl, alkoxy,        —C(═X)YR⁸, —YC(═X)R⁸, substituted or unsubstituted alkyl,        substituted or unsubstituted heteroalkyl, substituted or        unsubstituted aryl, substituted or unsubstituted alkyl-aryl,        substituted or unsubstituted cycloalkyl, substituted or        unsubstituted alkyl-cycloalkyl, substituted or unsubstituted        heteroaryl, substituted or unsubstituted alkyl-heteroaryl,        substituted or unsubstituted heterocycloalkyl, substituted or        unsubstituted alkyl-heterocycloalkyl, or -L-K;    -   R⁵ is H, hydroxy, alkyl, alkoxy, —C(═X)YR⁸, —YC(═X)R⁸,        substituted or unsubstituted alkyl, substituted or unsubstituted        heteroalkyl, substituted or unsubstituted aryl, substituted or        unsubstituted alkyl-aryl, substituted or unsubstituted        cycloalkyl, substituted or unsubstituted alkyl-cycloalkyl,        substituted or unsubstituted heteroaryl, substituted or        unsubstituted alkyl-heteroaryl, substituted or unsubstituted        heterocycloalkyl, substituted or unsubstituted        alkyl-heterocycloalkyl,    -   each R⁶, R⁷ is independently H, hydroxy, alkyl, alkoxy,        —C(═X)YR⁸, —YC(═X)R⁸, substituted or unsubstituted alkyl,        substituted or unsubstituted heteroalkyl, substituted or        unsubstituted aryl, substituted or unsubstituted alkyl-aryl,        substituted or unsubstituted cycloalkyl, substituted or        unsubstituted alkyl-cycloalkyl, substituted or unsubstituted        heteroaryl, substituted or unsubstituted alkyl-heteroaryl,        substituted or unsubstituted heterocycloalkyl, substituted or        unsubstituted alkyl-heterocycloalkyl, or -L-K; or R⁶ and R⁷        taken together form a bond;    -   each X is independently NH, S, or O;    -   each Y is independently NH, S, or O;    -   R⁸ is substituted or unsubstituted alkyl, substituted or        unsubstituted heteroalkyl, substituted or unsubstituted aryl,        substituted or unsubstituted alkyl-aryl, substituted or        unsubstituted cycloalkyl, substituted or unsubstituted        alkyl-cycloalkyl, substituted or unsubstituted heteroaryl,        substituted or unsubstituted alkyl-heteroaryl, substituted or        unsubstituted heterocycloalkyl, substituted or unsubstituted        alkyl-heterocycloalkyl, or -L-K;    -   L is A_(n), wherein        -   each A is independently NR¹, S(O)_(m), O, C(═X)Y, Y(C═X),            substituted or unsubstituted alkyl, substituted or            unsubstituted heteroalkyl, substituted or unsubstituted            aryl, substituted or unsubstituted heteroaryl, substituted            or unsubstituted cycloalkyl, or substituted or unsubstituted            heterocycloalkyl; wherein each m is independently 0-2;        -   n is 0-7;    -   K is a moiety that prevents systemic absorption;    -   provided that at least one of R¹, R², R³ or R⁴ is -L-K;

or a pharmaceutically acceptable prodrug thereof.

In some embodiments of a compound of Formula III, R¹ and R³ are -L-K. Insome embodiments, R¹, R² and R³ are -L-K.

In some embodiments, at least one of R¹, R², R³, R⁴, R⁵, R⁶ and R⁷ is H.In certain embodiments, R⁵, R⁶, R⁷ are H and R¹, R², R³ and R⁴ arealkyl, aryl, alkyl-aryl, or heteroalkyl. In some embodiments, R¹ and R²are H. In some embodiments, R¹, R², R⁵, R⁶ and R⁷ are H. In someembodiments, R⁶ and R⁷ together form a bond. In certain embodiments, R⁵,R⁶ and R⁷ are H, alkyl or O-alkyl.

In some embodiments, R¹ and R³ are -L-K. In some embodiments, R¹, R² andR³ are -L-K. In some embodiments, R³ and R⁴ are -L-K. In someembodiments, R¹ and R² together with the nitrogen to which they areattached form a 3-8 membered ring and the ring is substituted with -L-K.In some embodiments, R¹ or R² or R³ or R⁴ are aryl optionallysubstituted with -L-K. In some embodiments, R¹ or R² or R³ or R⁴ arealkyl optionally substituted with -L-K. In some embodiments, R¹ or R² orR³ or R⁴ are alky-aryl optionally substituted with -L-K. In someembodiments, R¹ or R² or R³ or R⁴ are heteroalkyl optionally substitutedwith -L-K.

In some embodiments, L is a C₁-C₇alkyl. In some embodiments, L isheteroalkyl. In certain embodiments, L is C₁-C₇alkyl-aryl. In someembodiments, L is C₁-C₇alkyl-aryl-C₁-C₇alkyl.

In certain embodiments, K is a non-protic charged group. In somespecific embodiments, each K is a ammonium group. In some embodiments,each K is a cyclic non-protic ammonium group. In some embodiments, eachK is an acyclic non-protic ammonium group.

In certain embodiments, each K is a cyclic non-protic ammonium group ofstructure:

In certain embodiments, K is an acyclic non-protic ammonium group ofstructure:

-   -   wherein p, q, R⁹, R¹⁰ and Z are as defined above. In certain        embodiments, p is 1. In other embodiments, p is 2. In further        embodiments, p is 3. In some embodiments, q is 0. In other        embodiments, q is 1. In some other embodiments, q is 2.

The compounds further comprise 1, 2, 3 or 4 anionic counterions selectedfrom Cl⁻, Br⁻, I⁻, R¹¹SO₃ ⁻, (SO₃ ⁻—R¹¹—SO₃ ⁻), R¹¹—CO₂ ⁻, (CO₂⁻—R¹¹—CO₂ ⁻), (R¹¹)₂(P═O)O⁻ and (R¹¹)(P═O)O₂ ²⁻ wherein R¹¹ is asdefined above. In some embodiments, the counterion is Cl⁻, Br⁻, I⁻,CH₂CO₂ ⁻, CH₃SO₃ ⁻, or C₆H₅SO₃ ⁻, or CO₂ ⁻—(CH₂)₂—CO₂ ⁻. In someembodiments, the compound of Formula III has one K group and onecounterion. In other embodiments, the compound of Formula III has one Kgroup, and two molecules of the compound of Formula III have onecounterion. In yet other embodiments, the compound of Formula III hastwo K groups and two counterions. In some other embodiments, thecompound of Formula III has one K group comprising two ammonium groupsand two counterions.

Also described herein are compounds having the Formula IIIA:

wherein:

-   -   each R¹, R² is independently H, substituted or unsubstituted        alkyl, or -L-K; or R¹ and R² together with the nitrogen to which        they are attached form a 3-8-membered ring that is optionally        substituted with R⁸;    -   and R³, R⁴, R⁸, L and K are as defined above.

In some embodiments of compounds of Formula IIIA, L is A_(n), whereineach A is substituted or unsubstituted alkyl, or substituted orunsubstituted heteroalkyl, and n is 0-7. In certain specific embodimentsof the compound of Formula IIIA, R¹ is H. In some embodiments of FormulaIIIA, R¹ and R² together with the nitrogen to which they are attachedform a 3-8-membered ring that is optionally substituted with -L-K.

Also described herein are compounds having the Formula IIIB:

wherein:

-   -   each R³, R⁴ is independently H, substituted or unsubstituted        alkyl, substituted or unsubstituted heteroalkyl, substituted or        unsubstituted aryl, substituted or unsubstituted alkyl-aryl, or        -L-K;    -   and R¹, R², L and K are as defined above.

In certain embodiments of Formula IIIB, R³ is H. In certain embodiments,R³ and R⁴ are each -L-K. In some embodiments, R³ is H and R⁴ issubstituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted aryl, substituted orunsubstituted alkyl-aryl containing one or two -L-K groups.

In some embodiments, an ASBTI suitable for the methods described hereinis a compound of Formula IIIC

wherein:

-   -   each R¹, R² is independently H, hydroxy, alkyl, alkoxy,        —C(═X)YR⁸, —YC(═X)R⁸, substituted or unsubstituted alkyl,        substituted or unsubstituted heteroalkyl, substituted or        unsubstituted aryl, substituted or unsubstituted alkyl-aryl,        substituted or unsubstituted cycloalkyl, substituted or        unsubstituted alkyl-cycloalkyl, substituted or unsubstituted        heteroaryl, substituted or unsubstituted alkyl-heteroaryl,        substituted or unsubstituted heterocycloalkyl, substituted or        unsubstituted alkyl-heterocycloalkyl, or -L-K; or R¹ and R²        together with the nitrogen to which they are attached form a        3-8-membered ring that is optionally substituted with R⁸;    -   each R³, R⁴ is independently H, hydroxy, alkyl, alkoxy,        —C(═X)YR⁸, —YC(═X)R⁸, substituted or unsubstituted alkyl,        substituted or unsubstituted heteroalkyl, substituted or        unsubstituted aryl, substituted or unsubstituted alkyl-aryl,        substituted or unsubstituted cycloalkyl, substituted or        unsubstituted alkyl-cycloalkyl, substituted or unsubstituted        heteroaryl, substituted or unsubstituted alkyl-heteroaryl,        substituted or unsubstituted heterocycloalkyl, substituted or        unsubstituted alkyl-heterocycloalkyl, or -L-K;    -   R⁵ is H, hydroxy, alkyl, alkoxy, —C(═X)YR⁸, —YC(═X)R⁸,        substituted or unsubstituted alkyl, substituted or unsubstituted        heteroalkyl, substituted or unsubstituted aryl, substituted or        unsubstituted alkyl-aryl, substituted or unsubstituted        cycloalkyl, substituted or unsubstituted alkyl-cycloalkyl,        substituted or unsubstituted heteroaryl, substituted or        unsubstituted alkyl-heteroaryl, substituted or unsubstituted        heterocycloalkyl, substituted or unsubstituted        alkyl-heterocycloalkyl,    -   each R⁶, R⁷ is independently H, hydroxy, alkyl, alkoxy,        —C(═X)YR⁸, —YC(═X)R⁸, substituted or unsubstituted alkyl,        substituted or unsubstituted heteroalkyl, substituted or        unsubstituted aryl, substituted or unsubstituted alkyl-aryl,        substituted or unsubstituted cycloalkyl, substituted or        unsubstituted alkyl-cycloalkyl, substituted or unsubstituted        heteroaryl, substituted or unsubstituted alkyl-heteroaryl,        substituted or unsubstituted heterocycloalkyl, substituted or        unsubstituted alkyl-heterocycloalkyl, or -L-K; or R⁶ and R⁷        taken together form a bond;    -   each X is independently NH, S, or O;    -   each Y is independently NH, S, or O;    -   R⁸ is substituted or unsubstituted alkyl, substituted or        unsubstituted heteroalkyl, substituted or unsubstituted aryl,        substituted or unsubstituted alkyl-aryl, substituted or        unsubstituted cycloalkyl, substituted or unsubstituted        alkyl-cycloalkyl, substituted or unsubstituted heteroaryl,        substituted or unsubstituted alkyl-heteroaryl, substituted or        unsubstituted heterocycloalkyl, substituted or unsubstituted        alkyl-heterocycloalkyl, or -L-K;    -   L is A_(n), wherein        -   each A is independently NR¹, S(O)_(m), O, C(═X)Y, Y(C═X),            substituted or unsubstituted alkyl, substituted or            unsubstituted heteroalkyl, substituted or unsubstituted            aryl, substituted or unsubstituted heteroaryl, substituted            or unsubstituted cycloalkyl, or substituted or unsubstituted            heterocycloalkyl; wherein each m is independently 0-2;        -   n is 0-7;    -   K is a moiety that prevents systemic absorption;        or a pharmaceutically acceptable salt thereof.

In some specific embodiments of Formula I, II or III, K is selected from

In some embodiments, an ASBTI suitable for the methods described hereinis a compound of Formula IV:

wherein

-   -   R¹ is a straight chain C₁₋₆ alkyl group;    -   R² is a straight chain C₁₋₆ alkyl group;    -   R³ is hydrogen or a group OR¹¹ in which R¹¹ is hydrogen,        optionally substituted C₁₋₆ alkyl or a C₁₋₆ alkylcarbonyl group;    -   R⁴ is pyridyl or an optionally substituted phenyl;    -   R⁵, R⁶ and R⁸ are the same or different and each is selected        from:        -   hydrogen, halogen, cyano, R¹⁵-acetylide, OR¹⁵, optionally            substituted C₁₋₆ alkyl, COR¹⁵, CH(OH)R¹⁵, S(O)_(n)R¹⁵,            P(O)(OR¹⁵)₂, OCOR⁵, OCF₃, OCN, SCN, NHCN, CH₂OR¹⁵, CHO,            (CH₂)_(p)CN, CONR¹²R¹³, (CH₂)_(p)CO₂R¹⁵, (CH₂)_(p)NR¹²R¹³,            CO₂R¹⁵, NHCOCF₃, NHSO₂R¹⁵, OCH₂OR¹⁵, OCH═CHR¹⁵,            O(CH₂CH₂O)_(n)R¹⁵, O(CH₂)_(p)SO₃R¹⁵, O(CH₂)_(p)NR¹²R¹³ and            O(CH₂)_(p)N⁺R¹²R¹³R¹⁴ wherein    -   p is an integer from 1-4,    -   n is an integer from 0-3 and    -   R¹², R¹³, R¹⁴ and R¹⁵ are independently selected from hydrogen        and optionally substituted C¹⁻⁶ alkyl;    -   R⁷ is a group of the formula

-   -   wherein the hydroxyl groups may be substituted by acetyl,        benzyl,        -   or —(C₁-C₆)-alkyl-R¹⁷,    -   wherein the alkyl group may be substituted with one or more        hydroxyl groups;

R¹⁶ is —COOH, —CH₂—OH, —CH₂—O-Acetyl, —COOMe or —COOEt;

R¹⁷ is H, —OH, —NH₂, —COOH or COOR¹⁸;

R¹⁸ is (C₁-C₄)-alkyl or —NH—(C₁-C₄)-alkyl;

X is —NH— or —O—; and

R⁹ and R¹⁰ are the same or different and each is hydrogen or C₁-C₆alkyl; and salts thereof.

In some embodiments, a compound of Formula IV has the structure ofFormula IVA or Formula IVB:

In some embodiments, a compound of Formula IV has the structure ofFormula IVC:

In some embodiments of Formula IV, X is O and R⁷ is selected from

In some embodiments, a compound of Formula IV is:

In some embodiments, an ASBTI suitable for the methods described hereinis a compound of Formula V:

wherein:

R¹¹ is selected from hydrogen or C₁₋₆alkyl;

One of R¹ and R² are selected from hydrogen or C₁₋₆alkyl and the otheris selected from C₁₋₆alkyl;

R^(x) and R^(y) are independently selected from hydrogen, hydroxy,amino, mercapto, C₁₋₆alkyl, C₁₋₆alkoxy, N—(C₁₋₆alkyl)amino,N,N—(C₁₋₆alkyl)₂amino, C₁₋₆alkylS(O)_(a) wherein a is 0 to 2;

R^(z) is selected from halo, nitr, cyano, hydroxy, amino, carboxy,carbamoyl, mercapto, sulphamoyl, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl,C₁₋₆alkoxy, C₁₋₆alkanoyl, C₁₋₆alkanoyloxy, N—(C₁₋₆alkyl)amino,N,N—(C₁₋₆alkyl)₂amino, C₁₋₆alkanoylamino, N—(C₁₋₆alkyl)carbamoyl,N,N—(C₁₋₆alkyl)₂carbamoyl, C₁₋₆alkylS(O)_(a) wherein a is 0 to 2,C₁₋₆alkoxycarbonyl, N—(C₁₋₆-alkyl)sulphamoyl andN,N—(C₁₋₆alkyl)₂sulphamoyl;

n is 0-5;

one of R⁴ and R⁵ is a group of formula (VA):

R³ and R⁶ and the other of R⁴ and R⁵ are independently selected fromhydrogen, halo, nitro, cyano, hydroxy, amino, carboxy, carbamoyl,mercapto, sulphamoyl, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₁₋₆alkoxy,C₁₋₆alkanoyl, C₁₋₆alkanoyloxy, N—(C₁₋₆alkyl)amino,N,N—(C₁₋₆alkyl)₂amino, C₁₋₆alkanoylamino, N—(C₁₋₆alkyl)carbamoyl,N,N—(C₁₋₆alkyl)₂carbamoyl, C₁₋₆alkylS(O)_(a) wherein a is 0 to 2,C₁₋₆alkoxycarbonyl, N—(C₁₋₆alkyl)sulphamoyl andN,N—(C₁₋₆alkyl)₂sulphamoyl;

-   -   wherein R³ and R⁶ and the other of R⁴ and R⁵ may be optionally        substituted on carbon by one or more R¹⁷;

X is —O—, —N(R^(a))—, —S(O)_(b)— or —CH(R^(a))—;

-   -   wherein R^(a) is hydrogen or C₁₋₆alkyl and b is 0-2;

Ring A is aryl or heteroaryl;

-   -   wherein Ring A is optionally substituted on carbon by one or        more substituents selected from R¹⁸;

R⁷ is hydrogen, C₁₋₆alkyl, carbocyclyl or heterocyclyl;

-   -   wherein R⁷ is optionally substituted on carbon by one or more        substituents selected from R¹⁹; and wherein if said heterocyclyl        contains an —NH— group, that nitrogen may be optionally        substituted by a group selected from R²⁰;

R⁸ is hydrogen or C₁₋₆-alkyl;

R⁹ is hydrogen or C₁₋₆alkyl;

R¹¹ is hydrogen, halo, nitro, cyano, hydroxy, amino, carbamoyl,mercapto, sulphamoyl, hydroxyaminocarbonyl, C₁₋₁₀alkyl, C₂₋₁₀alkynyl,C₂₋₁₀alkynyl, C₁₋₁₀alkoxy, C₁₋₁₀alkanoyl, C₁₋₁₀alkanoyloxy,N—(C₁₋₁₀alkyl)amino, N,N—(C₁₋₁₀alkyl)₂amino, N,N,N—(C₁₋₁₀alkyl)₃ammonio,C₁₋₁₀alkanoylamino, N—(C₁₋₁₀alkyl)carbamoyl, N,N—(C₁₋₁₀alkyl)₂carbamoyl,C₁₋₁₀alkylS(O)_(a) wherein a is 0 to 2, N—(C₁₋₁₀alkyl)sulphamoyl,N,N—(C₁₋₁₀alkyl)₂sulphamoyl, N—(C₁₋₁₀alkyl)sulphamoylamino,N,N—(C₁₋₁₀alkyl)₂sulphamoylamino, C₁₋₁₀alkoxycarbonylamino, carbocyclyl,carbocyclylC₁₋₁₀alkyl, heterocyclyl, heterocyclylC₁₋₁₀alkyl,carbocyclyl-(C₁₋₁₀alkylene)_(p)-R²¹—(C₁₋₁₀alkylene)_(q)- orheterocyclyl-(C₁₋₁₀alkylene), —R²²—(C₁₋₁₀alkylene)_(s)-; wherein R¹⁰ isoptionally substituted on carbon by one or more substituents selectedfrom R²³; and wherein if said heterocyclyl contains an —NH— group, thatnitrogen may be optionally substituted by a group selected from R²⁴; orR¹⁰ is a group of formula (VB):

wherein:

R¹¹ is hydrogen or C₁₋₆-alkyl;

R¹² and R¹³ are independently selected from hydrogen, halo, carbamoyl,sulphamoyl, C₁₋₁₀alkyl, C₂₋₁₀alkynyl, C₂₋₁₀alkynyl, C₁₋₁₀alkanoyl,N—(C₁₋₁₀alkyl)carbamoyl, N,N—(C₁₋₁₀alkyl)₂carbamoyl, C₁₋₁₀alkylS(O)_(a)wherein a is 0 to 2, N—(C₁₋₁₀alkyl)sulphamoyl,N,N—(C₁₋₁₀alkyl)₂sulphamoyl, N—(C₁₋₁₀alkyl)sulphamoylamino,N,N—(C₁₋₁₀alkyl)₂sulphamoylamino, carbocyclyl or heterocyclyl; whereinR¹² and R¹³ may be independently optionally substituted on carbon by oneor more substituents selected from R²⁵; and wherein if said heterocyclylcontains an —NH— group, that nitrogen may be optionally substituted by agroup selected from R²⁶;

R¹⁴ is selected from hydrogen, halo, carbamoyl, sulphamoyl,hydroxyaminocarbonyl, C₁₋₁₀alkyl, C₂₋₁₀alkenyl, C₂₋₁₀alkynyl,C₁₋₁₀alkanoyl, N—(C₁₋₁₀alkyl)carbamoyl, N,N—(C₁₋₁₀alkyl)₂carbamoyl,C₁₋₁₀alkylS(O)_(a) wherein a is 0 to 2, N—(C₁₋₁₀alkyl)sulphamoyl,N,N—(C₁₋₁₀alkyl)₂sulphamoyl, N—(C₁₋₁₀alkyl)sulphamoylamino,N,N—(C₁₋₁₀alkyl)₂sulphamoylamino, carbocyclyl, carbocyclylC₁₋₁₀alkyl,heterocyclyl, heterocyclylC₁₋₁₀alkyl,carbocyclyl-(C₁₋₁₀alkylene)_(p)-R²⁷—(C₁₋₁₀alkylene)_(q)- orheterocyclyl-(C₁₋₁₀alkylene)_(r)-R²⁸—(C₁₋₁₀alkylene)_(s)-; wherein R¹⁴may be optionally substituted on carbon by one or more substituentsselected from R²⁹; and wherein if said heterocyclyl contains an —NH—group, that nitrogen may be optionally substituted by a group selectedfrom R³⁰; or R¹⁴ is a group of formula (VC):

R¹⁵ is hydrogen or C₁₋₆alkyl; and R¹⁶ is hydrogen or C₁₋₆alkyl; whereinR¹⁶ may be optionally substituted on carbon by one or more groupsselected from R³¹;

or R¹⁵ and R¹⁶ together with the nitrogen to which they are attachedform a heterocyclyl; wherein said heterocyclyl may be optionallysubstituted on carbon by one or more R³⁷; and wherein if saidheterocyclyl contains an —NH— group, that nitrogen may be optionallysubstituted by a group selected from R³⁸;

m is 1-3; wherein the values of R⁷ may be the same or different;

R¹⁷, R¹⁸, R¹⁹, R²³, R²⁵, R²⁹, R³¹ and R³⁷ are independently selectedfrom halo, nitro, cyano, hydroxy, amino, carbamoyl, mercapto,sulphamoyl, hydroxyaminocarbonyl, C₁₋₁₀alkyl, C₂₋₁₀alkenyl,C₂₋₁₀alkynyl, C₁₋₁₀alkoxy, C₁₋₁₀alkanoyl, C₁₋₁₀alkanoyloxy,N—(C₁₋₁₀alkyl)amino, N,N—(C₁₋₁₀alkyl)₂amino, N,N,N—(C₁₋₁₀alkyl)₃ammonio,C₁₋₁₀alkanoylamino, N—(C₁₋₁₀alkyl)carbamoyl, N,N—(C₁₋₁₀alkyl)₂carbamoyl,C₁₋₁₀alkylS(O), wherein a is 0 to 2, N—(C₁₋₁₀alkyl)sulphamoyl,N,N—(C₁₋₁₀alkyl)₂sulphamoyl, N—(C₁₋₁₀alkyl)sulphamoylamino,N,N—(C₁₋₁₀alkyl)₂sulphamoylamino, C₁₋₁₀alkoxycarbonylamino, carbocyclyl,carbocyclylC₁₋₁₀alkyl, heterocyclyl, heterocyclylC₁₋₁₀alkyl,carbocyclyl-(C₁₋₁₀alkylene)_(p)-R³²—(C₁₋₁₀alkylene)_(q)- orheterocyclyl-(C₁₋₁₀alkylene)_(p)-R³³—(C₁₋₁₀alkylene)_(p)-; wherein R¹⁷,R¹⁸, R¹⁹, R²³, R²⁵, R²⁹, R³¹ and R³⁷ may be independently optionallysubstituted on carbon by one or more R³⁴; and wherein if saidheterocyclyl contains an —NH— group, that nitrogen may be optionallysubstituted by a group selected from R³⁵;

R²¹, R²², R²⁷, R²⁸, R³² or R³³ are independently selected from —O—,—NR³⁶—, —S(O)_(x)—, —NR³⁶C(O)NR³⁶—, —NR³⁶C(S)NR³⁶—, —OC(O)N═C—,—NR³⁶C(O)— or —C(O)NR³⁶—; wherein R³⁶ is selected from hydrogen orC₁₋₆alkyl, and x is 0-2;

p, q, r and s are independently selected from 0-2;

R³⁴ is selected from halo, hydroxy, cyano, carbamoyl, ureido, amino,nitro, carbamoyl, mercapto, sulphamoyl, trifluoromethyl,trifluoromethoxy, methyl, ethyl, methoxy, ethoxy, vinyl, allyl, ethynyl,formyl, acetyl, formamido, acetylamino, acetoxy, methylamino,dimethylamino, N-methylcarbamoyl, N,N-dimethylcarbamoyl, methylthio,methylsulphinyl, mesyl, N-methylsulphamoyl, N,N-dimethylsulphamoyl,N-methylsulphamoylamino and N,N-dimethylsulphamoylamino;

R²⁰, R²⁴, R²⁶, R³⁰, R³⁵ and R³⁸ are independently selected fromC₁₋₆alkyl, C₁₋₆alkanoyl, C₁₋₆alkylsulphonyl, C₁₋₆alkoxycarbonyl,carbamoyl, N—(C₁₋₆alkyl)carbamoyl, N,N—(C₁₋₆alkyl)carbamoyl, benzyl,benzyloxycarbonyl, benzoyl and phenylsulphonyl; and

wherein a “heteroaryl” is a totally unsaturated, mono or bicyclic ringcontaining 3-12 atoms of which at least one atom is chosen fromnitrogen, sulphur and oxygen, which heteroaryl may, unless otherwisespecified, be carbon or nitrogen linked;

wherein a “heterocyclyl” is a saturated, partially saturated orunsaturated, mono or bicyclic ring containing 3-12 atoms of which atleast one atom is chosen from nitrogen, sulphur and oxygen, whichheterocyclyl may, unless otherwise specified, be carbon or nitrogenlinked, wherein a —CH₂— group can optionally be replaced by a —C(O)—group, and a ring sulphur atom may be optionally oxidised to form anS-oxide; and

wherein a “carbocyclyl” is a saturated, partially saturated orunsaturated, mono or bicyclic carbon ring that contains 3-12 atoms;wherein a —CH₂— group can optionally be replaced by a —C(O) group; or apharmaceutically acceptable salt or in vivo hydrolysable ester or amideformed on an available carboxy or hydroxy group thereof.

In some embodiments, compound of Formula V is1,1-Dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N—[(R)-α-[N-(2-sulphoethyl)carbamoyl]-4-hydroxybenzyl]carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;1,1-Dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N—[(R)-α-[N—((S)-1-carboxy-2-(R)-hydroxypropyl)carbamoyl]-4-hydroxybenzyl]carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;or1,1-Dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N—[(R)-α-[N—((S)-1-carboxy-2-methylpropyl)carbamoyl]-4-hydroxybenzyl]carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine.

In some embodiments, compound of Formula V is

In some embodiments, an ASBTI suitable for the methods described hereinis a compound of Formula VI:

wherein:

R^(v) and R^(w) are independently selected from hydrogen or C₁₋₆alkyl;

one of R¹ and R² is selected from hydrogen or C₁₋₆alkyl and the other isselected from C₁₋₆alkyl;

R^(x) and R^(y) are independently selected from hydrogen or C₁₋₆alkyl,or one of R^(x) and R^(y) is hydrogen or C₁₋₆alkyl and the other ishydroxy or C₁₋₆alkoxy;

R^(z) is selected from halo, nitro, cyano, hydroxy, amino, carboxy,carbamoyl, mercapto, sulphamoyl, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl,C₁₋₆alkoxy, C₁₋₆alkanoyl, C₁₋₆alkanoyloxy, N—(C₁₋₆alkyl)amino,N,N—(C₁₋₆alkyl)₂amino, C₁₋₆alkanoylamino, N—(C₁₋₆alkyl)carbamoyl,N,N—(C₁₋₆alkyl)₂carbamoyl, C₁₋₆alkylS(O)_(a) wherein a is 0 to 2,C₁₋₆alkoxycarbonyl, N—(C₁₋₆alkyl)sulphamoyl andN,N—(C₁₋₆alkyl)₂sulphamoyl;

n is 0-5;

one of R⁴ and R⁵ is a group of formula (VIA):

R³ and R⁶ and the other of R⁴ and R⁵ are independently selected fromhydrogen, halo, nitro, cyano, hydroxy, amino, carboxy, carbamoyl,mercapto, sulphamoyl, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₁₋₆alkoxy,C₁₋₆alkanoyl, C₁₋₆alkanoyloxy, N—(C₁₋₆alkyl)amino,N,N—(C₁₋₆alkyl)₂amino, C₁₋₆alkanoylamino, N—(C₁₋₆alkyl)carbamoyl,N,N—(C₁₋₆alkyl)₂carbamoyl, C₁₋₆alkylS(O)_(a) wherein a is 0 to 2,C₁₋₆alkoxycarbonyl, N—(C₁₋₆alkyl)sulphamoyl andN,N—(C₁₋₆alkyl)₂sulphamoyl; wherein R³ and R⁶ and the other of R⁴ and R⁵may be optionally substituted on carbon by one or more R¹⁷;

X is —O—, —N(R^(a))—, —S(O)_(b)— or —CH(R^(a))—; wherein R^(a) ishydrogen or C₁₋₆alkyl and b is 0-2;

Ring A is aryl or heteroaryl; wherein Ring A is optionally substitutedon carbon by one or more substituents selected from R⁸;

R⁷ is hydrogen, C₁₋₆alkyl, carbocyclyl or heterocyclyl; wherein R⁷ isoptionally substituted on carbon by one or more substituents selectedfrom R¹⁹; and wherein if said heterocyclyl contains an —NH— group, thatnitrogen may be optionally substituted by a group selected from R²⁰;

R⁸ is hydrogen or C₁₋₆alkyl;

R⁹ is hydrogen or C₁₋₆alkyl;

R¹⁰ is hydrogen, halo, nitro, cyano, hydroxy, amino, carbamoyl,mercapto, sulphamoyl, hydroxyaminocarbonyl, C₁₋₁₀alkyl, C₂₋₁₀alkenyl,C₂₋₁₀alkynyl, C₁₋₁₀alkoxy, C₁₋₁₀alkanoyl, C₁₋₁₀alkanoyloxy,N—(C₁₋₁₀alkyl)amino, N,N—(C₁₋₁₀alkyl)₂amino, N,N,N—(C₁₋₁₀alkyl)₃ammonio,C₁₋₁₀alkanoylamino, N—(C₁₋₁₀alkyl)carbamoyl, N,N—(C₁₋₁₀alkyl)₂carbamoyl,C₁₋₁₀alkylS(O)_(a) wherein a is 0 to 2, N—(C₁₋₁₀alkyl)sulphamoyl,N,N—(C₁₋₁₀alkyl)₂sulphamoyl, N—(C₁₋₁₀alkyl)sulphamoylamino,N,N—(C₁₋₁₀alkyl)₂sulphamoylamino, C₁₋₁₀alkoxycarbonylamino, carbocyclyl,carbocyclylC₁₋₁₀alkyl, heterocyclyl, heterocyclylC₁₋₁₀alkyl,carbocyclyl-(C₁₋₁₀alkylene)_(p)-R²¹—(C₁₋₁₀alkylene)_(q)- orheterocyclyl-(C₁₋₁₀alkylene)_(r)-R²²—(C₁₋₁₀alkylene)_(s)-; wherein R¹⁰is optionally substituted on carbon by one or more substituents selectedfrom R²³; and wherein if said heterocyclyl contains an —NH— group, thatnitrogen may be optionally substituted by a group selected from R²⁴; orR¹⁰ is a group of formula (VIB):

wherein:

R¹¹ is hydrogen or C₁₋₆alkyl;

R¹² and R¹³ are independently selected from hydrogen, halo, nitro,cyano, hydroxy, amino, carbamoyl, mercapto, sulphamoyl, C₁₋₁₀alkyl,C₂₋₁₀alkenyl, C₂₋₁₀alkynyl, C₁₋₁₀alkoxy, C₁₋₁₀alkanoyl,C₁₋₁₀alkanoyloxy, N—(C₁₋₁₀alkyl)amino, N,N—(C₁₋₁₀alkyl)₂amino,C₁₋₁₀alkanoylamino, N—(C₁₋₁₀alkyl)carbamoyl, N,N—(C₁₋₁₀alkyl)₂carbamoyl,C₁₋₁₀alkylS(O)_(a) wherein a is 0 to 2, N—(C₁₋₁₀alkyl)sulphamoyl,N,N—(C₁₋₁₀alkyl)₂sulphamoyl, N—(C₁₋₁₀alkyl)sulphamoylamino,N,N—(C₁₋₁₀alkyl)₂sulphamoylamino, carbocyclyl or heterocyclyl; whereinR¹² and R¹³ may be independently optionally substituted on carbon by oneor more substituents selected from R²⁵; and wherein if said heterocyclylcontains an —NH— group, that nitrogen may be optionally substituted by agroup selected from R²⁶;

R¹⁴ is selected from hydrogen, halo, nitro, cyano, hydroxy, amino,carbamoyl, mercapto, sulphamoyl, hydroxyaminocarbonyl, C₁₋₁₀alkyl,C₂₋₁₀alkenyl, C₂₋₁₀alkynyl, C₁₋₁₀alkoxy, C₁₋₁₀alkanoyl,C₁₋₁₀alkanoyloxy, N—(C₁₋₁₀alkyl)amino, N,N—(C₁₋₁₀alkyl)₂amino,N,N,N—(C₁₋₁₀alkyl)₃ammonio, C₁₋₁₀alkanoylamino, N—(C₁₋₁₀alkyl)carbamoyl,N,N—(C₁₋₁₀alkyl)₂carbamoyl, C₁₋₁₀alkylS(O)_(a) wherein a is 0 to 2,N—(C₁₋₁₀alkyl)sulphamoyl, N,N—(C₁₋₁₀alkyl)₂sulphamoyl,N—(C₁₋₁₀alkyl)sulphamoylamino, N,N—(C₁₋₁₀alkyl)₂sulphamoylamino,C₁₋₁₀alkoxycarbonylamino, carbocyclyl, carbocyclylC₁₋₁₀alkyl,heterocyclyl, heterocyclylC₁₋₁₀alkyl,carbocyclyl-(C₁₋₁₀alkylene)_(p)-R²⁷—(C₁₋₁₀alkylene)_(q)- orheterocyclyl-(C₁₋₁₀alkylene)_(p)-R²⁸—(C₁₋₁₀alkylene)_(s)-; wherein R¹⁴may be optionally substituted on carbon by one or more substituentsselected from R²⁹; and wherein if said heterocyclyl contains an —NH—group, that nitrogen may be optionally substituted by a group selectedfrom R³⁰; or R¹⁴ is a group of formula (VIC):

R¹⁵ is hydrogen or C₁₋₆alkyl;

R¹⁶ is hydrogen or C₁₋₆alkyl; wherein R¹⁶ may be optionally substitutedon carbon by one or more groups selected from R³¹;

n is 1-3; wherein the values of R⁷ may be the same or different;

R¹⁷, R¹⁸, R¹⁹, R²³, R²⁵, R²⁹ or R³¹ are independently selected fromhalo, nitro, cyano, hydroxy, amino, carbamoyl, mercapto, sulphamoyl,hydroxyaminocarbonyl, amidino, C₁₋₁₀alkyl, C₂₋₁₀alkenyl, C₂₋₁₀alkynyl,C₁₋₁₀alkoxy, C₁₋₁₀alkanoyl, C₁₋₁₀alkanoyloxy, (C₁₋₁₀alkyl)₃silyl,N—(C₁₋₁₀alkyl)amino, N,N—(C₁₋₁₀alkyl)₂amino, N,N,N—(C₁₋₁₀alkyl)₃ammonio,C₁₋₁₀alkanoylamino, N—(C₁₋₁₀alkyl)carbamoyl, N,N—(C₁₋₁₀alkyl)₂carbamoyl,C₁₋₁₀alkylS(O)_(a) wherein a is 0 to 2, N—(C₁₋₁₀alkyl)sulphamoyl,N,N—(C₁₋₁₀alkyl)₂sulphamoyl, N—(C₁₋₁₀alkyl)sulphamoylamino,N,N—(C₁₋₁₀alkyl)₂sulphamoylamino, C₁₋₁₀alkoxycarbonylamino, carbocyclyl,carbocyclylC₁₋₁₀alkyl, heterocyclyl, heterocyclylC₁₋₁₀alkyl,carbocyclyl-(C₁₋₁₀alkylene)_(p)-R³²—(C₁₋₁₀alkylene)_(q)- orheterocyclyl-(C₁₋₁₀alkylene), —R³³—(C₁₋₁₀alkylene)_(s)-; wherein R¹⁷,R¹⁸, R¹⁹, R²³, R²⁵, R²⁹ or R³¹ may be independently optionallysubstituted on carbon by one or more R³⁴; and wherein if saidheterocyclyl contains an —NH— group, that nitrogen may be optionallysubstituted by a group selected from R³⁵;

R²¹, R²², R²⁷, R²⁸, R³² or R³³ are independently selected from —O—,—NR³⁶—, —S(O)_(x)—, NR³⁶C(O)NR³⁶—, —NR³⁶C(S)NR³⁶—, —OC(O)N═C—,—NR³⁶C(O)— or —C(O)NR³⁶—; wherein R³⁶ is selected from hydrogen orC₁₋₆alkyl, and x is 0-2;

p, q, r and s are independently selected from 0-2;

R³⁴ is selected from halo, hydroxy, cyano, carbamoyl, ureido, amino,nitro, carbamoyl, mercapto, sulphamoyl, trifluoromethyl,trifluoromethoxy, methyl, ethyl, methoxy, ethoxy, vinyl, allyl, ethynyl,formyl, acetyl, formamido, acetylamino, acetoxy, methylamino,dimethylamino, N-methylcarbamoyl, N,N-dimethylcarbamoyl, methylthio,methylsulphinyl, mesyl, N-methylsulphamoyl, N,N-dimethylsulphamoyl,N-methylsulphamoylamino and N,N-dimethylsulphamoylamino;

R²⁰, R²⁴, R²⁶, R³⁰ or R³⁵ are independently selected from C₁₋₆alkyl,C₁₋₆alkanoyl, C₁₋₆alkylsulphonyl, C₁₋₆alkoxycarbonyl, carbamoyl,N—(C₁₋₆alkyl)carbamoyl, N,N—(C₁₋₆alkyl)carbamoyl, benzyl,benzyloxycarbonyl, benzoyl and phenylsulphonyl;

or a pharmaceutically acceptable salt, solvate or solvate of such asalt, or an in vivo hydrolysable ester formed on an available carboxy orhydroxy thereof, or an in vivo hydrolysable amide formed on an availablecarboxy thereof.

In some embodiments, a compound of Formula VI has the structure ofFormula VID:

wherein:

R¹ and R² are independently selected from C₁₋₆alkyl; one of R⁴ and R⁵ isa group of formula (VIE):

R³ and R⁶ and the other of R⁴ and R⁵ are independently selected fromhydrogen, halo, nitro, cyano, hydroxy, amino, carboxy, carbamoyl,mercapto, sulphamoyl, C₁₋₄alkyl, C₂₋₄alkenyl, C₂₋₄alkynyl, C₁₋₄alkoxy,C₁₋₄alkanoyl, C₁₋₄alkanoyloxy, N—(C₁₋₄alkyl)amino,N,N—(C₁₋₄alkyl)₂amino, C₁₋₄alkanoylamino, N—(C₁₋₄alkyl)carbamoyl,N,N—(C₁₋₄alkyl)₂carbamoyl, C₁₋₄alkylS(O)_(a) wherein a is 0 to 2,C₁₋₄alkoxycarbonyl, N—(C₁₋₄alkyl)sulphamoyl andN,N—(C₁₋₄alkyl)₂sulphamoyl; wherein R³ and R⁶ and the other of R⁴ and R⁵may be optionally substituted on carbon by one or more R¹⁴;

R⁷ is carboxy, sulpho, sulphino, phosphono, —P(O)(OR^(a))(OR^(b)),P(O)(OH)(OR_(a)), —P(O)(OH)(R^(a)) or P(O)(OR^(a))(R^(b)), wherein R^(a)and R^(b) are independently selected from C₁₋₆alkyl; or R⁷ is a group offormula (VIF):

R⁸ and R⁹ are independently hydrogen, C₁₋₄alkyl or a saturated cyclicgroup, or R⁸ and R⁹ together form C₂₋₆alkylene; wherein R⁸ and R⁹ or R⁸and R⁹ together may be independently optionally substituted on carbon byone or more substituents selected from R¹⁵; and wherein if saidsaturated cyclic group contains an —NH— moiety, that nitrogen may beoptionally substituted by one or more R²⁰;

R¹⁰ is hydrogen or C₁₋₄alkyl; wherein R¹⁰ is optionally substituted oncarbon by one or more substituents selected from R²⁴;

R¹¹ is hydrogen, C₁₋₄alkyl, carbocyclyl or heterocyclyl; wherein R¹¹ isoptionally substituted on carbon by one or more substituents selectedfrom R¹⁶; and wherein if said heterocyclyl contains an —NH— moiety, thatnitrogen may be optionally substituted by one or more R²¹;

R¹² is hydrogen or C₁₋₄alkyl, carbocyclyl or heterocyclyl; wherein R¹²optionally substituted on carbon by one or more substituents selectedfrom R¹⁷; and wherein if said heterocyclyl contains an —NH— moiety, thatnitrogen may be optionally substituted by one or more R²²;

R¹³ is carboxy, sulpho, sulphino, phosphono, —P(O)(OR^(c))(OR^(d)),—P(O)(OH)(OR^(c)), —P(O)(OH)(R^(c)) or —P(O)(OR^(c))(R^(d)) whereinR^(c) and R^(d) are independently selected from C₁₋₆alkyl;

m is 1-3; wherein the values of R⁸ and R⁹ may be the same or different;

n is 1-3; wherein the values of R¹¹ may be the same or different;

p is 1-3; wherein the values of R¹² may be the same or different;

R¹⁴ and R¹⁶ are independently selected from halo, nitro, cyano, hydroxy,amino, carboxy, carbamoyl, mercapto, sulphamoyl, C₁₋₄alkyl, C₂₋₄alkenyl,C₂₋₄alkynyl, C₁₋₄alkoxy, C₁₋₄alkanoyl, C₁₋₄alkanoyloxy,N—(C₁₋₄alkyl)amino, N,N—(C₁₋₄alkyl)₂amino, C₁₋₄alkanoylamino,N—(C₁₋₄alkyl)carbamoyl, N,N—(C₁₋₄alkyl)₂carbamoyl, C₁₋₄alkylS(O)_(a)wherein a is 0 to 2, C₁₋₄alkoxycarbonyl, N—(C₁₋₄alkyl)sulphamoyl andN,N—(C₁₋₄alkyl)₂sulphamoyl; wherein R¹⁴ and R¹⁶ may be independentlyoptionally substituted on carbon by one or more R¹⁸;

R¹⁵ and R¹⁷ are independently selected from halo, nitro, cyano, hydroxy,amino, carboxy, carbamoyl, mercapto, sulphamoyl, C₁₋₄alkyl, C₂₋₄alkenyl,C₂₋₄alkynyl, C₁₋₄alkoxy, C₁₋₄alkanoyl, C₁₋₄alkanoyloxy,N—(C₁₋₄alkyl)amino, N,N—(C₁₋₄alkyl)₂amino, C₁₋₄alkanoylamino,N—(C₁₋₄alkyl)carbamoyl, N,N—(C₁₋₄alkyl)₂carbamoyl, C₁₋₄alkylS(O)_(a)wherein a is 0 to 2, C₁₋₄alkoxycarbonyl, N—(C₁₋₄alkyl)sulphamoyl andN,N—(C₁₋₄alkyl)₂sulphamoyl, carbocyclyl, heterocyclyl, sulpho, sulphino,amidino, phosphono, —P(O)(OR^(e))(OR^(f)), —P(O)(OH)(OR^(e)),—P(O)(OH)(R^(e)) or —P(O)(OR^(e))(R^(f)), wherein R^(e) and R^(f) areindependently selected from C₁₋₆alkyl; wherein R¹⁵ and R¹⁷ may beindependently optionally substituted on carbon by one or more R¹⁹; andwherein if said heterocyclyl contains an —NH— moiety, that nitrogen maybe optionally substituted by one or more R²³;

R¹⁸, R¹⁹ and R²⁵ are independently selected from halo, hydroxy, cyano,carbamoyl, ureido amino nitro, carboxy, carbamoyl, mercapto, sulphamoyl,trifluoromethyl, trifluoromethoxy, methyl, ethyl, methoxy, ethoxy,vinyl, allyl, ethynyl, methoxycarbonyl, formyl, acetyl, formamido,acetylamino, acetoxy, methylamino, dimethylamino, N-methylcarbamoyl,N,N-dimethylcarbamoyl, methylthio, methylsulphinyl, mesyl,N-methylsulphamoyl and N,N-dimethylsulphamoyl;

R²⁰, R²¹, R²², R²³ and R²⁶ are independently C₁₋₄alkyl, C₁₋₄alkanoyl,C₁₋₄alkylsulphonyl, sulphamoyl, N—(C₁₋₄alkyl)sulphamoyl,N,N—(C₁₋₄alkyl)₂sulphamoyl, C₁₋₄alkoxycarbonyl, carbamoyl,N—(C₁₋₄alkyl)carbamoyl, N,N—(C₁₋₄alkyl)₂carbamoyl, benzyl, phenethyl,benzoyl, phenylsulphonyl and phenyl;

R²⁴ is selected from halo, nitro, cyano, hydroxy, amino, carboxy,carbamoyl, mercapto, sulphamoyl, C₁₋₄alkyl, C₂₋₄alkenyl, C₂₋₄alkynyl,C₁₋₄alkoxy, C₁₋₄alkanoyl, C₁₋₄alkanoyloxy, N—(C₁₋₄alkyl)amino,N,N—(C₁₋₄alkyl)₂amino, C₁₋₄alkanoylamino, N—(C₁₋₄alkyl)carbamoyl,N,N—(C₁₋₄alkyl)₂carbamoyl, C₁₋₄alkylS(O)_(a) wherein a is 0 to 2,C₁₋₄alkoxycarbonyl, N—(C₁₋₄alkyl)sulphamoyl andN,N—(C₁₋₄alkyl)₂sulphamoyl, carbocyclyl, heterocyclyl; wherein R²⁴ maybe independently optionally substituted on carbon by one or more R²⁵;and wherein if said heterocyclyl contains an —NH— moiety, that nitrogenmay be optionally substituted by one or more R²⁶;

wherein any saturated cyclic group is a totally or partially saturated,mono or bicyclic ring containing 3-12 atoms of which 0-4 atoms arechosen from nitrogen, sulphur or oxygen, which may be carbon or nitrogenlinked;

wherein any heterocyclyl is a saturated, partially saturated orunsaturated, mono or bicyclic ring containing 3-12 atoms of which atleast one atom is chosen from nitrogen, sulphur or oxygen, which may becarbon or nitrogen linked, wherein a —CH₂— group can optionally bereplaced by a —C(O)— or a ring sulphur atom may be optionally oxidisedto form the S-oxides; and

wherein any carbocyclyl is a saturated, partially saturated orunsaturated, mono or bicyclic carbon ring that contains 3-12 atoms,wherein a —CH₂— group can optionally be replaced by a —C(O)—;

or a pharmaceutically acceptable salt thereof.

In some embodiments, any compound described herein is covalentlyconjugated to a bile acid using any suitable method. In someembodiments, compounds described herein are covalently bonded to acyclodextrin or a biodegradable polymer (e.g., a polysaccharide).

In certain embodiments compounds described herein are not systemicallyabsorbed. Moreover, provided herein are compounds that inhibit bile saltrecycling in the gastrointestinal tract of an individual. In someembodiments, compounds described herein, may not be transported from thegut lumen and/or do not interact with ASBT. In some embodiments,compounds described herein, do not affect, or minimally affect, fatdigestion and/or absorption. In certain embodiments, the administrationof a therapeutically effective amount of any compound described hereindoes not result in gastrointestinal disturbance or lactic acidosis in anindividual. In certain embodiments, compounds described herein areadministered orally. In some embodiments, an ASBTI is released in thedistal ileum. An ASBTI compatible with the methods described herein maybe a direct inhibitor, an allosteric inhibitor, or a partial inhibitorof the Apical Sodium-dependent Bile acid Transporter.

In certain embodiments, compounds that inhibit ASBT or any recuperativebile acid transporters are compounds that are described in EP1810689,U.S. Pat. Nos. 6,458,851, 7,413,536, 7,514,421, US Appl. PublicationNos. 2002/0147184, 2003/0119809, 2003/0149010, 2004/0014806,2004/0092500, 2004/0180861, 2004/0180860, 2005/0031651, 2006/0069080,2006/0199797, 2006/0241121, 2007/0065428, 2007/0066644, 2007/0161578,2007/0197628, 2007/0203183, 2007/0254952, 2008/0070888, 2008/0070892,2008/0070889, 2008/0070984, 2008/0089858, 2008/0096921, 2008/0161400,2008/0167356, 2008/0194598, 2008/0255202, 2008/0261990, WO 2002/50027,WO2005/046797, WO2006/017257, WO2006/105913, WO2006/105912,WO2006/116499, WO2006/117076, WO2006/121861, WO2006/122186,WO2006/124713, WO2007/050628, WO2007/101531, WO2007/134862,WO2007/140934, WO2007/140894, WO2008/028590, WO2008/033431,WO2008/033464, WO2008/031501, WO2008/031500, WO2008/033465,WO2008/034534, WO2008/039829, WO2008/064788, WO2008/064789,WO2008/088836, WO2008/104306, WO2008/124505, and WO2008/130616; thecompounds described therein that inhibit recuperative bile acidtransport are hereby incorporated herein by reference.

In certain embodiments, compounds that inhibit ASBT or any recuperativebile acid transporters are compounds described in WO93/16055,WO94/18183, WO94/18184, WO96/05188, WO96/08484, WO96/16051, WO97/33882,WO98/38182, WO99/35135, WO98/40375, WO99/64409, WO99/64410, WO00/01687,WO00/47568, WO00/61568, DE 19825804, WO00/38725, WO00/38726, WO00/38727(including those compounds with a 2,3,4,5-tetrahydro-1-benzothiepine1,1-dioxide structure), WO00/38728, WO01/66533, WO02/50051, EP0864582(e.g.(3R,5R)-3-butyl-3-ethyl-1,1-dioxido-5-Phenyl-2,3,4,5-tetrahydro-1,4-benzo-thiazepin-8-yl(□-D-glucopyranosiduronic acid, WO94/24087, WO98/07749, WO98/56757,WO99/32478, WO99/35135, WO00/20392, WO00/20393, WO00/20410, WO00/20437,WO01/34570, WO00/35889, WO01/68637, WO01/68096, WO02/08211, WO3/020710,WO03/022825, WO03/022830, WO03/0222861, JP10072371, U.S. Pat. Nos.5,910,494; 5,723,458; 5,817,652; 5,663,165; 5,998,400; 6,465,451,5,994,391; 6,107,494; 6,387,924; 6,784,201; 6,875,877; 6,740,663;6,852,753; 5,070,103, 6,114,322, 6,020,330, 7,179,792, EP251315,EP417725, EP489-423, EP549967, EP573848, EP624593, EP624594, EP624595,EP869121, EP1070703, WO04/005247, compounds disclosed as having IBATactivity in Drugs of the Future, 24, 425-430 (1999), Journal ofMedicinal Chemistry, 48, 5837-5852, (2005) and Current MedicinalChemistry, 13, 997-1016, (2006); the compounds described therein thatinhibit recuperative bile acid transport are hereby incorporated hereinby reference.

In some embodiments, compounds that inhibit ASBT or any recuperativebile acid transporter are benzothiepines, benzothiazepines (including1,2-benzothiazepines; 1,4-benzothiazepines; 1,5-benzothiazepines; and/or1,2,5-benzothiadiazepines). In some embodiments, compounds that inhibitASBT or any recuperative bile acid transporter include and are notlimited to S-8921 (disclosed in EP597107, WO 93/08155), 264W94 (GSK)disclosed in WO 96/05188; SC-435(1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethanesulfonate salt), SC-635 (Searle); 2164U90(3-butyl-3-ethyl-2,3,4,5-tetrahydro-5-phenyl-1,4-benzothiazepine1,1-dioxide); BARI-1741 (Aventis SA), AZD 7508 (Astra Zeneca); barixibat(11-(D-gluconamido)-N-{2-[(1S,2R,3S)-3-hydroxy-3-phenyl-2-(2-pyridyl)-1-(2-pyridylamino)propyl]phenyl}undecanamide)or the like, or combinations thereof. In some embodiments, an ASBTI is:

In certain embodiments, compounds described herein have one or morechiral centers. As such, all stereoisomers are envisioned herein. Invarious embodiments, compounds described herein are present in opticallyactive or racemic forms. It is to be understood that the compounds ofthe present invention encompasses racemic, optically-active,regioisomeric and stereoisomeric forms, or combinations thereof thatpossess the therapeutically useful properties described herein.Preparation of optically active forms is achieve in any suitable manner,including by way of non-limiting example, by resolution of the racemicform by recrystallization techniques, by synthesis from optically-activestarting materials, by chiral synthesis, or by chromatographicseparation using a chiral stationary phase. In some embodiments,mixtures of one or more isomer is utilized as the therapeutic compounddescribed herein. In certain embodiments, compounds described hereincontains one or more chiral centers. These compounds are prepared by anymeans, including enantioselective synthesis and/or separation of amixture of enantiomers and/or diastereomers. Resolutions there ofcompounds and isomers thereof is achieved by any means including, by wayof non-limiting example, chemical processes, enzymatic processes,fractional crystallization, distillation, chromatography, and the like.

The compounds described herein, and other related compounds havingdifferent substituents are synthesized using techniques and materialsdescribed herein and as described, for example, in Fieser and Fieser'sReagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons,1991); Rodd's Chemistry of Carbon Compounds, Volumes 1-5 andSupplementals (Elsevier Science Publishers, 1989); Organic Reactions,Volumes 1-40 (John Wiley and Sons, 1991), Larock's Comprehensive OrganicTransformations (VCH Publishers Inc., 1989), March, DVANCED ORGANICCHEMISTRY 4^(th) Ed., (Wiley 1992); Carey and Sundberg, ADVANCED ORGANICCHEMISTRY 4′ Ed., Vols. A and B (Plenum 2000, 2001), and Green and Wuts,PROTECTIVE GROUPS IN ORGANIC SYNTHESIS 3^(rd) Ed., (Wiley 1999) (all ofwhich are incorporated by reference for such disclosure). Generalmethods for the preparation of compound as described herein are modifiedby the use of appropriate reagents and conditions, for the introductionof the various moieties found in the formulae as provided herein. As aguide the following synthetic methods are utilized.

Formation of Covalent Linkages by Reaction of an Electrophile with aNucleophile

The compounds described herein are modified using various electrophilesand/or nucleophiles to form new functional groups or substituents. TableA entitled “Examples of Covalent Linkages and Precursors Thereof” listsselected non-limiting examples of covalent linkages and precursorfunctional groups which yield the covalent linkages. Table A is used asguidance toward the variety of electrophiles and nucleophilescombinations available that provide covalent linkages. Precursorfunctional groups are shown as electrophilic groups and nucleophilicgroups.

TABLE A Examples of Covalent Linkages and Precursors Thereof CovalentLinkage Product Electrophile Nucleophile Carboxamides Activated estersamines/anilines Carboxamides acyl azides amines/anilines Carboxamidesacyl halides amines/anilines Esters acyl halides alcohols/phenols Estersacyl nitriles alcohols/phenols Carboxamides acyl nitrilesamines/anilines Imines Aldehydes amines/anilines Hydrazones aldehydes orketones Hydrazines Oximes aldehydes or ketones Hydroxylamines Alkylamines alkyl halides amines/anilines Esters alkyl halides carboxylicacids Thioethers alkyl halides Thiols Ethers alkyl halidesalcohols/phenols Thioethers alkyl sulfonates Thiols Esters alkylsulfonates carboxylic acids Ethers alkyl sulfonates alcohols/phenolsEsters Anhydrides alcohols/phenols Carboxamides Anhydridesamines/anilines Thiophenols aryl halides Thiols Aryl amines aryl halidesAmines Thioethers Azindines Thiols Boronate esters Boronates GlycolsCarboxamides carboxylic acids amines/anilines Esters carboxylic acidsAlcohols hydrazines Hydrazides carboxylic acids N-acylureas orAnhydrides carbodiimides carboxylic acids Esters diazoalkanes carboxylicacids Thioethers Epoxides Thiols Thioethers haloacetamides ThiolsAmmotriazines halotriazines amines/anilines Triazinyl ethershalotriazines alcohols/phenols Amidines imido esters amines/anilinesUreas Isocyanates amines/anilines Urethanes Isocyanates alcohols/phenolsThioureas isothiocyanates amines/anilines Thioethers Maleimides ThiolsPhosphite esters phosphoramidites Alcohols Silyl ethers silyl halidesAlcohols Alkyl amines sulfonate esters amines/anilines Thioetherssulfonate esters Thiols Esters sulfonate esters carboxylic acids Etherssulfonate esters Alcohols Sulfonamides sulfonyl halides amines/anilinesSulfonate esters sulfonyl halides phenols/alcoholsUse of Protecting Groups

In the reactions described, it is necessary to protect reactivefunctional groups, for example hydroxy, amino, imino, thio or carboxygroups, where these are desired in the final product, in order to avoidtheir unwanted participation in reactions. Protecting groups are used toblock some or all of the reactive moieties and prevent such groups fromparticipating in chemical reactions until the protective group isremoved. In some embodiments it is contemplated that each protectivegroup be removable by a different means. Protective groups that arecleaved under totally disparate reaction conditions fulfill therequirement of differential removal.

In some embodiments, protective groups are removed by acid, base,reducing conditions (such as, for example, hydrogenolysis), and/oroxidative conditions. Groups such as trityl, dimethoxytrityl, acetal andt-butyldimethylsilyl are acid labile and are used to protect carboxy andhydroxy reactive moieties in the presence of amino groups protected withCbz groups, which are removable by hydrogenolysis, and Fmoc groups,which are base labile. Carboxylic acid and hydroxy reactive moieties areblocked with base labile groups such as, but not limited to, methyl,ethyl, and acetyl in the presence of amines blocked with acid labilegroups such as t-butyl carbamate or with carbamates that are both acidand base stable but hydrolytically removable.

In some embodiments carboxylic acid and hydroxy reactive moieties areblocked with hydrolytically removable protective groups such as thebenzyl group, while amine groups capable of hydrogen bonding with acidsare blocked with base labile groups such as Fmoc. Carboxylic acidreactive moieties are protected by conversion to simple ester compoundsas exemplified herein, which include conversion to alkyl esters, or areblocked with oxidatively-removable protective groups such as2,4-dimethoxybenzyl, while co-existing amino groups are blocked withfluoride labile silyl carbamates.

Allyl blocking groups are useful in then presence of acid- andbase-protecting groups since the former are stable and are subsequentlyremoved by metal or pi-acid catalysts. For example, an allyl-blockedcarboxylic acid is deprotected with a Pd⁰-catalyzed reaction in thepresence of acid labile t-butyl carbamate or base-labile acetate amineprotecting groups. Yet another form of protecting group is a resin towhich a compound or intermediate is attached. As long as the residue isattached to the resin, that functional group is blocked and does notreact. Once released from the resin, the functional group is availableto react.

Typically blocking/protecting groups are selected from:

Other protecting groups, plus a detailed description of techniquesapplicable to the creation of protecting groups and their removal aredescribed in Greene and Wuts, Protective Groups in Organic Synthesis,3rd Ed., John Wiley & Sons, New York, N.Y., 1999, and Kocienski,Protective Groups, Thieme Verlag, New York, N.Y., 1994, which areincorporated herein by reference for such disclosure.

In some embodiments, ASBTIs described herein are synthesized asdescribed in, for example, WO 96/05188, U.S. Pat. Nos. 5,994,391;7,238,684; 6,906,058; 6,020,330; and 6,114,322. In some embodiments,ASBTIs described herein are synthesized starting from compounds that areavailable from commercial sources or that are prepared using proceduresoutlined herein. In some embodiments, compounds described herein areprepared according to the process set forth in Scheme 1:

In certain embodiments, the synthesis begins with a reaction of1,4-diazabicyclo[2.2.2]octane with 4-iodo-1-chloro butane to provide acompound of structure 1-I. Such compounds are prepared in any suitablemanner, e.g., as set forth in Tremont, S. J. et. al., J. Med. Chem.2005, 48, 5837-5852. The compound of structure 1-I is then subjected toa reaction with phenethylamine to provide a compound of structure 1-II.The compound of structure 1-II is then allowed to react withdicyanodiamide to provide a compound of Formula I.

In some embodiments, a first compound of Formula III is subjected to afurther reaction to provide a second compound of Formula III as shown inScheme 2 below.

A first compound of Formula III, 1-IA, is alkylated with iodomethane toprovide a second compound of Formula III, 1-IB. Alkylation of 1-IB witha compound of structure 2-II provides a further compound of Formula III,IC. In an alternative embodiment, a first compound of Formula III, 1-IA,is alkylated with a compound of structure 2-I to provide a secondcompound of Formula III, 1-IC.

General Definitions

The term “bile acid,” as used herein, includes steroid acids (and/or thecarboxylate anion thereof), and salts thereof, found in the bile of ananimal (e.g., a human), including, by way of non-limiting example,cholic acid, cholate, deoxycholic acid, deoxycholate, hyodeoxycholicacid, hyodeoxycholate, glycocholic acid, glycocholate, taurocholic acid,taurocholate, chenodeoxycholic acid, chenodeoxycholate, lithocholicacid, lithocolate, and the like. Taurocholic acid and/or taurocholateare referred to herein as TCA. Any reference to a bile acid used hereinincludes reference to a bile acid, one and only one bile acid, one ormore bile acids, or to at least one bile acid. Therefore, the terms“bile acid,” “bile salt,” “bile acid/salt,” “bile acids,” “bile salts,”and “bile acids/salts” are, unless otherwise indicated, utilizedinterchangeably herein. Any reference to a bile acid used hereinincludes reference to a bile acid or a salt thereof. Furthermore,pharmaceutically acceptable bile acid esters are optionally utilized asthe “bile acids” described herein, e.g., bile, acids conjugated to anamino acid (e.g., glycine or taurine). Other bile acid esters include,e.g., substituted or unsubstituted alkyl ester, substituted orunsubstituted heteroalkyl esters, substituted or unsubstituted arylesters, substituted or unsubstituted heteroaryl esters, or the like. Forexample, the term “bile acid” includes cholic acid conjugated witheither glycine or taurine: glycocholate and taurocholate, respectively(and salts thereof). Any reference to a bile acid used herein includesreference to an identical compound naturally or synthetically prepared.Furthermore, it is to be understood that any singular reference to acomponent (bile acid or otherwise) used herein includes reference to oneand only one, one or more, or at least one of such components.Similarly, any plural reference to a component used herein includesreference to one and only one, one or more, or at least one of suchcomponents, unless otherwise noted. Moreover, as used herein, bileacid/salt mimics or mimetics described herein are compounds that mimicthe agonist signaling properties of the bile acid/salt, especially atTGR5 (GPBAR1, BG37, Axor109) receptors. Examples includes thosedescribed in WO 2010/014836, which is incorporated herein for suchdisclosure. In some embodiments, bile acid mimetics includetriterpenoid, such as oleanoic acid, ursolic acid, or the like.

The term “subject”, “patient” or “individual” are used interchangeablyherein and refer to mammals and non-mammals, e.g., suffering from adisorder described herein. Examples of mammals include, but are notlimited to, any member of the mammalian class: humans, non-humanprimates such as chimpanzees, and other apes and monkey species; farmanimals such as cattle, horses, sheep, goats, swine; domestic animalssuch as rabbits, dogs, and cats; laboratory animals including rodents,such as rats, mice and guinea pigs, and the like. Examples ofnon-mammals include, but are not limited to, birds, fish and the like.In one embodiment of the methods and compositions provided herein, themammal is a human.

The term “colon,” as used herein, includes the cecum, ascending colon,hepatic flexure, splenic flexure, descending colon, and sigmoid.

The term “composition,” as used herein includes the disclosure of both acomposition and a composition administered in a method as describedherein. Furthermore, in some embodiments, the composition of the presentinvention is or comprises a “formulation,” an oral dosage form or arectal dosage form as described herein.

The terms “treat,” “treating” or “treatment,” and other grammaticalequivalents as used herein, include alleviating, inhibiting or reducingsymptoms, reducing or inhibiting severity of, reducing incidence of,prophylactic treatment of, reducing or inhibiting recurrence of,preventing, delaying onset of, delaying recurrence of, abating orameliorating a disease or condition symptoms, ameliorating theunderlying metabolic causes of symptoms, inhibiting the disease orcondition, e.g., arresting the development of the disease or condition,relieving the disease or condition, causing regression of the disease orcondition, relieving a condition caused by the disease or condition, orstopping the symptoms of the disease or condition. The terms furtherinclude achieving a therapeutic benefit. By therapeutic benefit is meanteradication or amelioration of the underlying disorder being treated,and/or the eradication or amelioration of one or more of thephysiological symptoms associated with the underlying disorder such thatan improvement is observed in the patient.

The terms “prevent,” “preventing” or “prevention,” and other grammaticalequivalents as used herein, include preventing additional symptoms,preventing the underlying metabolic causes of symptoms, inhibiting thedisease or condition, e.g., arresting the development of the disease orcondition and are intended to include prophylaxis. The terms furtherinclude achieving a prophylactic benefit. For prophylactic benefit, thecompositions are optionally administered to a patient at risk ofdeveloping a particular disease, to a patient reporting one or more ofthe physiological symptoms of a disease, or to a patient at risk ofreoccurrence of the disease.

Where combination treatments or prevention methods are contemplated, itis not intended that the agents described herein be limited by theparticular nature of the combination. For example, the agents describedherein are optionally administered in combination as simple mixtures aswell as chemical hybrids. An example of the latter is where the agent iscovalently linked to a targeting carrier or to an active pharmaceutical.Covalent binding can be accomplished in many ways, such as, though notlimited to, the use of a commercially available cross-linking agent.Furthermore, combination treatments are optionally administeredseparately or concomitantly.

As used herein, the terms “pharmaceutical combination”, “administeringan additional therapy”, “administering an additional therapeutic agent”and the like refer to a pharmaceutical therapy resulting from the mixingor combining of more than one active ingredient and includes both fixedand non-fixed combinations of the active ingredients. The term “fixedcombination” means that at least one of the agents described herein, andat least one co-agent, are both administered to a patient simultaneouslyin the form of a single entity or dosage. The term “non-fixedcombination” means that at least one of the agents described herein, andat least one co-agent, are administered to a patient as separateentities either simultaneously, concurrently or sequentially withvariable intervening time limits, wherein such administration provideseffective levels of the two or more agents in the body of the patient.In some instances, the co-agent is administered once or for a period oftime, after which the agent is administered once or over a period oftime. In other instances, the co-agent is administered for a period oftime, after which, a therapy involving the administration of both theco-agent and the agent are administered. In still other embodiments, theagent is administered once or over a period of time, after which, theco-agent is administered once or over a period of time. These also applyto cocktail therapies, e.g. the administration of three or more activeingredients.

As used herein, the terms “co-administration”, “administered incombination with” and their grammatical equivalents are meant toencompass administration of the selected therapeutic agents to a singlepatient, and are intended to include treatment regimens in which theagents are administered by the same or different route of administrationor at the same or different times. In some embodiments the agentsdescribed herein will be co-administered with other agents. These termsencompass administration of two or more agents to an animal so that bothagents and/or their metabolites are present in the animal at the sametime. They include simultaneous administration in separate compositions,administration at different times in separate compositions, and/oradministration in a composition in which both agents are present. Thus,in some embodiments, the agents described herein and the other agent(s)are administered in a single composition. In some embodiments, theagents described herein and the other agent(s) are admixed in thecomposition.

The terms “effective amount” or “therapeutically effective amount” asused herein, refer to a sufficient amount of at least one agent beingadministered which achieve a desired result, e.g., to relieve to someextent one or more symptoms of a disease or condition being treated. Incertain instances, the result is a reduction and/or alleviation of thesigns, symptoms, or causes of a disease, or any other desired alterationof a biological system. In certain instances, an “effective amount” fortherapeutic uses is the amount of the composition comprising an agent asset forth herein required to provide a clinically significant decreasein a disease. An appropriate “effective” amount in any individual caseis determined using any suitable technique, such as a dose escalationstudy.

The terms “administer,” “administering”, “administration,” and the like,as used herein, refer to the methods that may be used to enable deliveryof agents or compositions to the desired site of biological action.These methods include, but are not limited to oral routes, intraduodenalroutes, parenteral injection (including intravenous, subcutaneous,intraperitoneal, intramuscular, intravascular or infusion), topical andrectal administration. Administration techniques that are optionallyemployed with the agents and methods described herein are found insources e.g., Goodman and Gilman, The Pharmacological Basis ofTherapeutics, current ed.; Pergamon; and Remington's, PharmaceuticalSciences (current edition), Mack Publishing Co., Easton, Pa. In certainembodiments, the agents and compositions described herein areadministered orally.

The term “pharmaceutically acceptable” as used herein, refers to amaterial that does not abrogate the biological activity or properties ofthe agents described herein, and is relatively nontoxic (i.e., thetoxicity of the material significantly outweighs the benefit of thematerial). In some instances, a pharmaceutically acceptable material maybe administered to an individual without causing significant undesirablebiological effects or significantly interacting in a deleterious mannerwith any of the components of the composition in which it is contained.

The term “carrier” as used herein, refers to relatively nontoxicchemical agents that, in certain instances, facilitate the incorporationof an agent into cells or tissues.

The term “non-systemic” or “minimally absorbed” as used herein refers tolow systemic bioavailability and/or absorption of an administeredcompound. In some instances a non-systemic compound is a compound thatis substantially not absorbed systemically. In some embodiments, ASBTIcompositions described herein deliver the ASBTI to the distal ileum,colon, and/or rectum and not systemically (e.g., a substantial portionof the ASBTI is not systemically absorbed. In some embodiments, thesystemic absorption of a non-systemic compound is <0.1%, <0.3%, <0.5%,<0.6%, <0.7%, <0.8%, <0.9%, <1%, <1.5%, <2%, <3%, or <5% of theadministered dose (wt. % or mol %). In some embodiments, the systemicabsorption of a non-systemic compound is <15% of the administered dose.In some embodiments, the systemic absorption of a non-systemic compoundis <25% of the administered dose. In an alternative approach, anon-systemic ASBTI is a compound that has lower systemic bioavailabilityrelative to the systemic bioavailability of a systemic ASBTI (e.g.,compound 100A, 100C). In some embodiments, the bioavailability of anon-systemic ASBTI described herein is <30%, <40%, <50%, <60%, or <70%of the bioavailability of a systemic ASBTI (e.g., compound 100A, 100C).

In another alternative approach, the compositions described herein areformulated to deliver <10% of the administered dose of the ASBTIsystemically. In some embodiments, the compositions described herein areformulated to deliver <20% of the administered dose of the ASBTIsystemically. In some embodiments, the compositions described herein areformulated to deliver <30% of the administered dose of the ASBTIsystemically. In some embodiments, the compositions described herein areformulated to deliver <40% of the administered dose of the ASBTIsystemically. In some embodiments, the compositions described herein areformulated to deliver <50% of the administered dose of the ASBTIsystemically. In some embodiments, the compositions described herein areformulated to deliver <60% of the administered dose of the ASBTIsystemically. In some embodiments, the compositions described herein areformulated to deliver <70% of the administered dose of the ASBTIsystemically. In some embodiments, systemic absorption is determined inany suitable manner, including the total circulating amount, the amountcleared after administration, or the like.

The term “ASBT inhibitor” refers to a compound that inhibits apicalsodium-dependent bile transport or any recuperative bile salt transport.The term Apical Sodium-dependent Bile Transporter (ASBT) is usedinterchangeably with the term Ileal Bile Acid Transporter (IBAT).

The term “reducing food intake” refers to consumption of a lower amountof food by an individual undergoing therapy with any ASBTI describedherein compared to the amount of food consumed in the absence of ASBTItherapy.

The term “induction of satiety” or “inducing satiety” or “satiety”refers to a feeling of fullness and/or a reduction of the sensation ofhunger.

The term “metabolic disorder” refers to any disorder that involves analteration in the normal metabolism of carbohydrates, lipids, proteins,nucleic acids or a combination thereof. A metabolic disorder isassociated with either a deficiency or excess in a metabolic pathwayresulting in an imbalance in metabolism of nucleic acids, proteins,lipids, and/or carbohydrates. Factors affecting metabolism include, andare not limited to, the endocrine (hormonal) control system (e.g., theinsulin pathway, the enteroendocrine hormones including GLP-1, GLP-2,oxyntomodulin, PYY or the like), the neural control system (e.g., GLP-1in the brain) or the like. Examples of metabolic disorders include andare not limited to diabetes, insulin resistance, dyslipidemia, metabolicsyndrome, or the like.

The term “enhancing enteroendocrine peptide secretion” refers to asufficient increase in the level of the enteroendocrine peptide agentto, for example, decrease hunger in a subject, to curb appetite in asubject and/or decrease the food intake of a subject or individualand/or treat any disease or disorder described herein. In someembodiments, enhanced enteroendocrine peptide secretion reverses oralleviates symptoms of congestive heart failure, ventriculardysfunction, toxic hypervolemia, polycystic ovary syndrome, inflammatorybowel disease, impaired bowel integrity, short bowel syndrome,gastritis, peptic ulcer, or irritable bowel syndrome.

In various embodiments, pharmaceutically acceptable salts describedherein include, by way of non-limiting example, a nitrate, chloride,bromide, phosphate, sulfate, acetate, hexafluorophosphate, citrate,gluconate, benzoate, propionate, butyrate, sulfosalicylate, maleate,laurate, malate, fumarate, succinate, tartrate, amsonate, pamoate,p-tolunenesulfonate, mesylate and the like. Furthermore,pharmaceutically acceptable salts include, by way of non-limitingexample, alkaline earth metal salts (e.g., calcium or magnesium), alkalimetal salts (e.g., sodium-dependent or potassium), ammonium salts andthe like.

The term “optionally substituted” or “substituted” means that thereferenced group substituted with one or more additional group(s). Incertain embodiments, the one or more additional group(s) areindividually and independently selected from amide, ester, alkyl,cycloalkyl, heteroalkyl, aryl, heteroaryl, heteroalicyclic, hydroxy,alkoxy, aryloxy, alkylthio, arylthio, alkylsulfoxide, arylsulfoxide,ester, alkylsulfone, arylsulfone, cyano, halo, alkoyl, alkoyloxo,isocyanato, thiocyanato, isothiocyanato, nitro, haloalkyl, haloalkoxy,fluoroalkyl, amino, alkyl-amino, dialkyl-amino, amido.

An “alkyl” group refers to an aliphatic hydrocarbon group. Reference toan alkyl group includes “saturated alkyl” and/or “unsaturated alkyl”.The alkyl group, whether saturated or unsaturated, includes branched,straight chain, or cyclic groups. By way of example only, alkyl includesmethyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, sec-butyl,t-butyl, pentyl, iso-pentyl, neo-pentyl, and hexyl. In some embodiments,alkyl groups include, but are in no way limited to, methyl, ethyl,propyl, isopropyl, butyl, isobutyl, tertiary butyl, pentyl, hexyl,ethenyl, propenyl, butenyl, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, and the like. A “lower alkyl” is a C₁-C₆ alkyl. A“heteroalkyl” group substitutes any one of the carbons of the alkylgroup with a heteroatom having the appropriate number of hydrogen atomsattached (e.g., a CH₂ group to an NH group or an O group).

An “alkoxy” group refers to a (alkyl)O— group, where alkyl is as definedherein.

The term “alkylamine” refers to the —N(alkyl)_(x)H_(y) group, whereinalkyl is as defined herein and x and y are selected from the group x=1,y=1 and x=2, y=0. When x=2, the alkyl groups, taken together with thenitrogen to which they are attached, optionally form a cyclic ringsystem.

An “amide” is a chemical moiety with formula —C(O)NHR or —NHC(O)R, whereR is selected from alkyl, cycloalkyl, aryl, heteroaryl (bonded through aring carbon) and heteroalicyclic (bonded through a ring carbon).

The term “ester” refers to a chemical moiety with formula —C(═O)OR,where R is selected from the group consisting of alkyl, cycloalkyl,aryl, heteroaryl and heteroalicyclic.

As used herein, the term “aryl” refers to an aromatic ring wherein eachof the atoms forming the ring is a carbon atom. Aryl rings describedherein include rings having five, six, seven, eight, nine, or more thannine carbon atoms. Aryl groups are optionally substituted. Examples ofaryl groups include, but are not limited to phenyl, and naphthalenyl.

The term “cycloalkyl” refers to a monocyclic or polycyclic non-aromaticradical, wherein each of the atoms forming the ring (i.e. skeletalatoms) is a carbon atom. In various embodiments, cycloalkyls aresaturated, or partially unsaturated. In some embodiments, cycloalkylsare fused with an aromatic ring. Cycloalkyl groups include groups havingfrom 3 to 10 ring atoms. Illustrative examples of cycloalkyl groupsinclude, but are not limited to, the following moieties:

and the like. Monocyclic cycloalkyls include, but are not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, andcyclooctyl.

The term “heterocyclo” refers to heteroaromatic and heteroalicyclicgroups containing one to four ring heteroatoms each selected from O, Sand N. In certain instances, each heterocyclic group has from 4 to 10atoms in its ring system, and with the proviso that the ring of saidgroup does not contain two adjacent O or S atoms. Non-aromaticheterocyclic groups include groups having 3 atoms in their ring system,but aromatic heterocyclic groups must have at least 5 atoms in theirring system. The heterocyclic groups include benzo-fused ring systems.An example of a 3-membered heterocyclic group is aziridinyl (derivedfrom aziridine). An example of a 4-membered heterocyclic group isazetidinyl (derived from azetidine). An example of a 5-memberedheterocyclic group is thiazolyl. An example of a 6-membered heterocyclicgroup is pyridyl, and an example of a 10-membered heterocyclic group isquinolinyl. Examples of non-aromatic heterocyclic groups arepyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl,tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino,morpholino, thiomorpholino, thioxanyl, piperazinyl, aziridinyl,azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl,oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl,2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl,1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl,dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl,imidazolidinyl, 3-azabicyclo[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl,3H-indolyl and quinolizinyl. Examples of aromatic heterocyclic groupsare pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl,tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl,isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl,benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl,phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl,oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl,benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl,naphthyridinyl, and furopyridinyl.

The terms “heteroaryl” or, alternatively, “heteroaromatic” refers to anaryl group that includes one or more ring heteroatoms selected fromnitrogen, oxygen and sulfur. An N-containing “heteroaromatic” or“heteroaryl” moiety refers to an aromatic group in which at least one ofthe skeletal atoms of the ring is a nitrogen atom. In certainembodiments, heteroaryl groups are monocyclic or polycyclic.Illustrative examples of heteroaryl groups include the followingmoieties:

and the like.

A “heteroalicyclic” group or “heterocyclo” group refers to a cycloalkylgroup, wherein at least one skeletal ring atom is a heteroatom selectedfrom nitrogen, oxygen and sulfur. In various embodiments, the radicalsare with an aryl or heteroaryl. Illustrative examples of heterocyclogroups, also referred to as non-aromatic heterocycles, include:

and the like. The term heteroalicyclic also includes all ring forms ofthe carbohydrates, including but not limited to the monosaccharides, thedisaccharides and the oligosaccharides.

The term “halo” or, alternatively, “halogen” means fluoro, chloro, bromoand iodo.

The terms “haloalkyl,” and “haloalkoxy” include alkyl and alkoxystructures that are substituted with one or more halogens. Inembodiments, where more than one halogen is included in the group, thehalogens are the same or they are different. The terms “fluoroalkyl” and“fluoroalkoxy” include haloalkyl and haloalkoxy groups, respectively, inwhich the halo is fluorine.

The term “heteroalkyl” include optionally substituted alkyl, alkenyl andalkynyl radicals which have one or more skeletal chain atoms selectedfrom an atom other than carbon, e.g., oxygen, nitrogen, sulfur,phosphorus, silicon, or combinations thereof. In certain embodiments,the heteroatom(s) is placed at any interior position of the heteroalkylgroup. Examples include, but are not limited to, —CH₂—O—CH₃,—CH₂—CH₂—O—CH₃, —CH₂—NH—CH₃, —CH₂—CH₂—NH—CH₃, —CH₂—N(CH₃)—CH₃,—CH₂—CH₂—NH—CH₃, —CH₂—CH₂—N(CH₃)—CH₃, —CH₂—S—CH₂—CH₃,—CH₂—CH₂,—S(O)—CH₃, —CH₂—CH₂—S(O)₂—CH₃, —CH═CH—O—CH₃, —Si(CH₃)₃,—CH₂—CH═N—OCH₃, and —CH═CH—N(CH₃)—CH₃. In some embodiments, up to twoheteroatoms are consecutive, such as, by way of example, —CH₂—NH—OCH₃and —CH₂—O—Si(CH₃)₃.

A “cyano” group refers to a —CN group.

An “isocyanato” group refers to a —NCO group.

A “thiocyanato” group refers to a —CNS group.

An “isothiocyanato” group refers to a —NCS group.

“Alkoyloxy” refers to a RC(═O)O— group.

“Alkoyl” refers to a RC(═O)— group.

The term “modulate,” as used herein refers to having some affect on(e.g., increasing, enhancing or maintaining a certain level).

The term “optionally substituted” or “substituted” means that thereferenced group may be substituted with one or more additional group(s)individually and independently selected from C₁-C₆alkyl,C₃-C₈cycloalkyl, aryl, heteroaryl, C₂-C₆heteroalicyclic, hydroxy,C₁-C₆alkoxy, aryloxy, C₁-C₆alkylthio, arylthio, C₁-C₆alkylsulfoxide,arylsulfoxide, C₁-C₆alkylsulfone, arylsulfone, cyano, halo, C₂-C₅acyl,C₂-C₈acyloxy, nitro, C₁-C₆haloalkyl, C₁-C₆fluoroalkyl, and amino,including C₁-C₆alkylamino, and the protected derivatives thereof. By wayof example, an optional substituents may be L^(s)R^(s), wherein eachL^(s) is independently selected from a bond, —O—, —C(═O)—, —S—, —S(═O)—,—S(═O)₂—, —NH—, —NHC(═O)—, —C(═O)NH—, S(═O)₂NH—, —NHS(═O)₂—, —OC(═O)NH—,—NHC(═O)O—, —(C₁-C₆alkyl)-, or —(C₂-C₆alkenyl)-; and each R^(s) isindependently selected from H, (C₁-C₄alkyl), (C₃-C₅cycloalkyl),heteroaryl, aryl, and C₁-C₆heteroalkyl. Optionally substitutednon-aromatic groups may be substituted with one or more oxo (═O). Theprotecting groups that may form the protective derivatives of the abovesubstituents are known to those of skill in the art and may be found inreferences such as Greene and Wuts, above. In some embodiments, alkylgroups described herein are optionally substituted with an O that isconnected to two adjacent carbon atoms (i.e., forming an epoxide).

The term “therapeutically effective amount” or an “effective amount” asused herein, refers to a sufficient amount of a therapeutically activeagent to provide a desired effect in a subject or individual. In someembodiments, a “therapeutically effective amount” or an “effectiveamount” of an enteroendocrine peptide secretion enhancing agent refersto a sufficient amount of the enteroendocrine peptide secretionenhancing agent to a treat a metabolic disorder in a subject orindividual. In some embodiments, a “therapeutically effective amount” oran “effective amount” of an enteroendocrine peptide secretion enhancingagent refers to a sufficient amount of the enteroendocrine peptidesecretion enhancing agent to increase the secretion of enteroendocrinepeptide(s) in a subject or individual. In specific embodiments, the“therapeutically effective amount” is an amount that when delivered tothe colon or rectum it produces an anorectal response (e.g., it increasethe secretion of enteroendocrine peptide(s) in the rectum and/or colon,particularly in the L-cells, of an individual). In some embodiments, a“therapeutically effective amount” or an “effective amount” of anenteroendocrine peptide secretion enhancing agent refers to a sufficientamount of the enteroendocrine peptide secretion enhancing agent todecrease hunger in a subject, to curb appetite in a subject and/ordecrease the food intake of a subject or individual.

L-Cells

L-cells are scattered throughout the epithelial layer of the gut fromthe duodenum to the rectum, with the highest numbers occurring in theileum, colon, and rectum. They are characterized by an open-cellmorphology, with apical microvilli facing into the gut lumen andsecretory vesicles located adjacent to the basolateral membrane, and aretherefore in direct contact with nutrients in the intestinal lumen.Furthermore, L-cells are located in close proximity to both neurons andthe microvasculature of the intestine, thereby allowing the L-cell to beaffected by both neural and hormonal signals. As well as Glucagon-LikePeptide 1 (GLP-1) and Glucagon-Like Peptide 2 (GLP-2), L-cells alsosecrete the anorexigenic hormone, peptide YY (PYY), and glutamate. Thecells are just one member of a much larger family of enteroendocrinecells that secrete a range of hormones, including ghrelin, GIP,cholecystokinin, somatostatin, and secretin, which are involved in thelocal coordination of gut physiology, as well as in playing wider rolesin the control of insulin release and appetite. L-cells are unevenlydistributed in the gastrointestinal tract, within higher concentrationsin the distal portion of the gastrointestinal tract (e.g., in the distalileum, colon and rectum).

Proglucagon Products

The proglucagon gene product is expressed in the L-cells of the smallintestine, in □-cells of the pancreas and in the central nervous system.Tissue-specific expression of isoforms of the enzyme prohormoneconvertase directs posttranslational synthesis of specificproglucagon-derived peptides in the L-cell and α-cell. Specifically,cleavage of proglucagon by prohormone convertase 1/3, which is expressedin the L-cell, forms GLP-1 and GLP-2, as well as the glucagon-containingpeptides, glicentin and oxyntomodulin. In contrast, α-cell expression ofprohormone convertase 2 forms glucagon, glicentin-related pancreaticpeptide, and the major proglucagon fragment, which contains within itssequence both the GLP-1 and GLP-2 sequences.

Glucagon-like peptide 1 (GLP-1) is an intestinal hormone that effects inthe regulation of glycemia, stimulating glucose-dependent insulinsecretion, proinsulin gene expression, and ß-cell proliferative andanti-apoptotic pathways, as well as inhibiting glucagon release, gastricemptying, and food intake. The anorexigenic effect of GLP-1 is mediatedby GLP-1 receptors which are present in both the NTS and hypothalamus,and in the pancreas, lung, brain, kidney, gastrointestinal tract andheart. Reduced secretion of GLP-1 contributes to the pathogenesis ofobesity and enhanced/or normal secretion restores satiety.

The primary physiological stimulus of GLP-1 secretion from L-cells isingestion of carbohydrates, luminal glucose (not systemic glucose) fat,and protein. Protein hydrolysate are also potent triggers of GLP-1release, and certain amino acids such as, but not limited to, alanine,serine, glutamine, asparagine, and glycine stimulate GLP-1 release.Within the fat group, the long-chain unsaturated fatty acid andshort-chain fatty acid subgroups are potent triggers of GLP-1 release,while the short-chain fatty acids also stimulate peptide YY release. Inaddition to luminal nutrients, intestinal peptides, neurotransmitters,as well as systemic hormones, modulate GLP-1 secretion. Such intestinalpeptides include, but are not limited to, somatostatin (forms SS14 andSS28), and such neurotransmitters include, but are not limited to,acetylcholine and □-aminobutyric acid (GABA) (both of which enhanceGLP-1 release), and □- and □-adrenergic agonists, (which respectivelyinhibit and/or stimulate GLP-1 secretion from L-cells). Peripheralhormones that participate in energy homeostasis, such as the adipocytehormone leptin, also stimulate GLP-1 release. Other GLP-1 secretegouesinclude bile acids/salts, insulin, gastrin-releasing peptide (GRP),several gut peptides including, but not limited to, Gastric InhibitoryPolypeptide (GIP) and calcitonin gene-related protein (CGRP). CGRP is apeptide found throughout the enteric nervous system. Thus, GLP-1secretagogues include, but are not limited to, nutrients,neurotransmitters, neuropeptides, intestinal peptide, peripheralhormones, and bile acis/salts.

Within about 15 minutes of food ingestion the circulating GLP-1 levelsincrease and remain elevated for up to 3 hours, depending on thecomposition of the meal. Circulating GLP-1 exists in two equipotentforms, GLP-1^(7-36NH2) and GLP-1⁷⁻³⁷, with GLP-1^(7-36NH2) being thepredominant form. Secreted GLP-1 is rapidly degraded by the ubiquitousenzyme dipeptidyl peptidase-4 (DPP-4), resulting in an extremely shorthalf-life for GLP-1 of about 30 seconds to about 2 minutes. Therefore,levels of circulating GLP-1 are maintained by inhibiting DPP-4 activity,or alternatively, by enhancing GLP-1 secretion.

In some embodiments, provided herein is a method of increasingcirculating GLP (e.g., GLP-1) levels by administering to the distalgastrointestinal tract (e.g., distal ileum, colon and/or rectum) aneffective amount of an enteroendocrine peptide secretion enhancing agent(e.g., a bile acid). In certain embodiments, the method is included in amethod of treating a metabolic disorder. In some embodiments, the methodis included in a method of treating obesity or diabetes. FIG. 19illustrates the increase of circulating GLP-1 levels following rectaladministration of an enteroendocrine peptide secretion enhancing agent(e.g., a bile acid). FIG. 19 illustrates the increase of circulatingGLP-1 levels following rectal administration of taurocholic acid.

Pancreatic Polypeptide (PP)-Fold Peptides

The Pancreatic Polypeptide (PP)-fold peptides include Peptide YY (PYY),Pancreatic Polypeptide (PP) and Neuropeptide Y (NPY), which all sharesequence homology and contain several tyrosine residues. They have acommon tertiary structure which consists of an □-helix and polyprolinehelix, connected by a ß-turn, resulting in a characteristic U-shapedpeptide, the PP-fold.

Neuropeptide Y (NPY) is one of the most abundant neurotransmitters inthe brain. Hypothalamic levels of NPY reflect the body's nutritionalstatus, wherein the levels of hypothalamic NPY mRNA and NPY releaseincrease with fasting and decrease after feeding.

Pancreatic Polypeptide (PP) is produced by cells at the periphery of theislets of the endocrine pancreas, and to a lesser extent in the exocrinepancreas, colon and rectum. The release of PP is biphasic and occurs inproportion to the number of calories ingested, with the levels remainingelevated for up to 6 hours post-prandially. The circulating levels of PPare increased by gastric distension, ghrelin, motilin and secretin andreduced by somatostatin. In addition, circulating PP exhibits a diurnalrhythm, with levels low in the early hours of the morning and highest inthe evening. The levels of PP have been found to reflect long-termenergy stores, with lower levels and reduced second phase of release inobese subjects, and higher levels in anorexic subjects. Circulating PPis unable to cross the blood-brain barrier, but exerts its anorecticeffect by sending anorectic signals via brainstem pathways, hypothalamicneuropeptides and by modulating expression of other gut hormones such asghrelin.

Peptide YY (PYY) is secreted predominantly from the distalgastrointestinal tract, particularly the ileum, colon and rectum. FIG.18 illustrates the concentration of PYY at various locations in thegastrointestinal tract. The L-cells of the intestine release PYY inproportion to the amount of calories ingested, and occurs before thenutrients reach the cells in the distal tract. Thus, release may bemediated via a neural reflex as well as direct contact with nutrients.Post-prandially, the circulating PYY levels rise rapidly to a plateauafter 1-2 hours and remain elevated for up to 6 hours. The levels of PYYare also influenced by meal composition, with higher levels obtainedfollowing fat intake relative to carbohydrate or protein intake. Othersignals, such as gastric acid, CCK and luminal bile salts, insulin-likegrowth factor 1, bombesin and calcitonin-gene-related peptide increasePYY levels, whereas gastric distension has no effect, and levels arereduced by GLP-1. The N-terminal of circulating PYY allows it to crossthe blood-brain barrier.

In some embodiments, provided herein is a method of increasingcirculating PYY levels by administering to the distal gastrointestinaltract (e.g., distal ileum, colon and/or rectum) an effective amount ofan enteroendocrine peptide secretion enhancing agent (e.g., a bileacid). In certain embodiments, the method is included in a method oftreating a metabolic disorder. In some embodiments, the method isincluded in a method of treating obesity or diabetes. FIG. 20illustrates the increase of circulating PYY levels following rectaladministration of an enteroendocrine peptide secretion enhancing agent(e.g., a bile acid).

Enteroendocrine Peptide Secretion Enhanced Treatment

Intravenous administration of GLP-1 decreases food intake in both leanand obese individuals in a dose-dependent manner, although the effect issmall when infusions achieve post-prandial circulating levels. Inaddition, obese subjects given subcutaneous GLP-1 prior to each meal,reduce their caloric intake. Thus, modulation/control of GLP-1 secretionrestore satiety and controls appetite and food intake. Similarly,infusion of PP decreases appetite and food intake. Obese humans have arelatively low level of circulating PYY and a relative deficiency ofpost-prandial secretion of PYY. Administration of PYY causes a delay ingastric emptying, a delay in secretions from the pancreas and stomach,and increases the absorption of fluids and electrolytes from the ileumafter a meal. In addition, intravenous administration of PYY decreasesappetite, inhibits food intake and reduces weight gain.

Appetite, weight gain and food intake are controlled by the circulatingor systemic levels of GLP-1, PYY and PP. The methods and compositiondescribed herein use, by way of non-limiting example, the administrationof bile acids/salts and bile acids/salts mimics to modulate (e.g.,increase) the circulating levels of GLP-1, PYY and PP. In certainembodiments of the present invention, such administration decreases foodintake and weight gain while suppressing appetite.

Bile Acid

Bile contains water, electrolytes and a numerous organic moleculesincluding bile acids, cholesterol, phospholipids and bilirubin. Bile issecreted from the liver and stored in the gall bladder, and upon gallbladder contraction, due to ingestion of a fatty meal, bile passesthrough the bile duct into the intestine. Bile acids are critical fordigestion and absorption of fats and fat-soluble vitamins in the smallintestine. Adult humans produce 400 to 800 mL of bile daily. Thesecretion of bile can be considered to occur in two stages. Initially,hepatocytes secrete bile into canaliculi, from which it flows into bileducts and this hepatic bile contains large quantities of bile acids,cholesterol and other organic molecules. Then, as bile flows through thebile ducts, it is modified by addition of a watery, bicarbonate-richsecretion from ductal epithelial cells. Bile is concentrated, typicallyfive-fold, during storage in the gall bladder.

The flow of bile is lowest during fasting, and a majority of that isdiverted into the gallbladder for concentration. When chyme from aningested meal enters the small intestine, acid and partially digestedfats and proteins stimulate secretion of cholecystokinin and secretin,both of which are important for secretion and flow of bile.Cholecystokinin (cholecysto=gallbladder and kinin=movement) is a hormonewhich stimulates contractions of the gallbladder and common bile duct,resulting in delivery of bile into the gut. The most potent stimulus forrelease of cholecystokinin is the presence of fat in the duodenum.Secretin is a hormone secreted in response to acid in the duodenum, andit simulates biliary duct cells to secrete bicarbonate and water, whichexpands the volume of bile and increases its flow out into theintestine.

Bile acids are derivatives of cholesterol. Cholesterol, ingested as partof the diet or derived from hepatic synthesis, are converted into bileacids in the hepatocyte. Examples of such bile acids include cholic andchenodeoxycholic acids, which are then conjugated to an amino acid (suchas glycine or taurine) to yield the conjugated form that is activelysecreted into cannaliculi. The most abundant of the bile salts in humansare cholate and deoxycholate, and they are normally conjugated witheither glycine or taurine to give glycocholate or taurocholaterespectively.

Free cholesterol is virtually insoluble in aqueous solutions, however inbile it is made soluble by the presence of bile acids and lipids.Hepatic synthesis of bile acids accounts for the majority of cholesterolbreakdown in the body. In humans, roughly 500 mg of cholesterol areconverted to bile acids and eliminated in bile every day. Therefore,secretion into bile is a major route for elimination of cholesterol.Large amounts of bile acids are secreted into the intestine every day,but only relatively small quantities are lost from the body. This isbecause approximately 95% of the bile acids delivered to the duodenumare absorbed back into blood within the ileum, by a process is known as“Enterohepatic Recirculation”.

Venous blood from the ileum goes straight into the portal vein, andhence through the sinusoids of the liver. Hepatocytes extract bile acidsvery efficiently from sinusoidal blood, and little escapes the healthyliver into systemic circulation. Bile acids are then transported acrossthe hepatocytes to be resecreted into canaliculi. The net effect of thisenterohepatic recirculation is that each bile salt molecule is reusedabout 20 times, often two or three times during a single digestivephase. Bile biosynthesis represents the major metabolic fate ofcholesterol, accounting for more than half of the approximate 800 mg/dayof cholesterol that an average adult uses up in metabolic processes. Incomparison, steroid hormone biosynthesis consumes only about 50 mg ofcholesterol per day. Much more that 400 mg of bile salts is required andsecreted into the intestine per day, and this is achieved by re-cyclingthe bile salts. Most of the bile salts secreted into the upper region ofthe small intestine are absorbed along with the dietary lipids that theyemulsified at the lower end of the small intestine. They are separatedfrom the dietary lipid and returned to the liver for re-use.R^(e)-cycling thus enables 20-30 g of bile salts to be secreted into thesmall intestine each day.

Bile acids are amphipathic, with the cholesterol-derived portioncontaining both hydrophobic (lipid soluble) and polar (hydrophilic)moieties while the amino acid conjugate is generally polar andhydrophilic. This amphipathic nature enables bile acids to carry out twoimportant functions: emulsification of lipid aggregates andsolubilization and transport of lipids in an aqueous environment. Bileacids have detergent action on particles of dietary fat which causes fatglobules to break down or to be emulsified. Emulsification is importantsince it greatly increases the surface area of fat available fordigestion by lipases which cannot access the inside of lipid droplets.Furthermore, bile acids are lipid carriers and are able to solubilizemany lipids by forming micelles and are critical for transport andabsorption of the fat-soluble vitamins.

Pharmaceutical Compositions and Methods of Use

In some embodiments, compositions described herein are administered fordelivery of enteroendocrine peptide secretion enhancing agents to asubject or individual. In certain embodiments, any compositionsdescribed herein are formulated for ileal, rectal and/or colonicdelivery. In more specific embodiments, the composition is formulatedfor non-systemic or local delivery to the rectum and/or colon. It is tobe understood that as used herein, delivery to the colon includesdelivery to sigmoid colon, transverse colon, and/or ascending colon. Instill more specific embodiments, the composition is formulated fornon-systemic or local delivery to the rectum and/or colon isadministered rectally. In other specific embodiments, the composition isformulated for non-systemic or local delivery to the rectum and/or colonis administered orally.

In some embodiments, provided herein is a composition comprising anenteroendocrine peptide secretion enhancing agent and, optionally, apharmaceutically acceptable carrier for reducing food intake in anindividual. In some embodiments, provided herein is a compositioncomprising an enteroendocrine peptide secretion enhancing agent and,optionally, a pharmaceutically acceptable carrier for reducingcirculating glucose levels in an individual. In some embodiments,provided herein is a composition comprising an enteroendocrine peptidesecretion enhancing agent and, optionally, a pharmaceutically acceptablecarrier for increasing insulin levels in an individual. In specificembodiments, the composition is formulated for delivering theenteroendocrine peptide secretion enhancing agent to the distalgastrointestinal tract of the individual. Generally, a therapeuticallyeffective amount of an enteroendocrine peptide secretion enhancing agentis provided.

In certain embodiments, the composition comprises an enteroendocrinepeptide secretion enhancing agent and an absorption inhibitor. Inspecific embodiments, the absorption inhibitor is an inhibitor thatinhibits the absorption of the (or at least one of the) specificenteroendocrine peptide secretion enhancing agent with which it iscombined. In some embodiments, the composition comprises anenteroendocrine peptide secretion enhancing agent, an absorptioninhibitor and a carrier (e.g., an orally suitable carrier or a rectallysuitable carrier, depending on the mode of intended administration). Incertain embodiments, the composition comprises an enteroendocrinepeptide secretion enhancing agent, an absorption inhibitor, a carrier,and one or more of a cholesterol absorption inhibitor, anenteroendocrine peptide, a peptidase inhibitor, a spreading agent, and awetting agent.

In certain embodiments enteroendocrine peptide-secretion enhancingagents are selected from, by way of non-limiting example, bile acids,bile acid mimic and/or modified bile acids. In more specificembodiments, compositions described herein are formulated fornon-systemic or local delivery of a bile acid, bile acid mimic and/ormodified bile acid (as the active component or components) to the rectumand/or colon, including the sigmoid colon, transverse colon, and/orascending colon. In certain embodiments, the compositions describedherein are administered rectally for non-systemic or local delivery ofthe bile acid active component to the rectum and/or colon, including thesigmoid colon, transverse colon, and/or ascending colon. In otherembodiments, the compositions described herein are administered orallyfor non-systemic delivery of the bile salt active component to therectum and/or colon, including the sigmoid colon, transverse colon,and/or ascending colon. In specific embodiments, compositions formulatedfor oral administration are, by way of non-limiting example, entericallycoated or formulated oral dosage forms, such as, tablets and/orcapsules. It is to be understood that the terms “subject” and“individual” are utilized interchangeably herein and include, e.g.,humans and human patients in need of treatment.

Enteroendocrine Peptide Enhancing Agents

In some embodiments, enteroendocrine peptide enhancing agents providedherein include, by way of non-limiting example, enteroendocrine peptidesecretion (e.g., of the L-cells) enhancing agents, inhibitors ofdegradation of enteroendocrine peptides (e.g., of the L-cells), orcombinations thereof.

In certain embodiments, the enteroendocrine peptide secretion enhancingagents used in the methods and compositions described herein include, byway of non-limiting example, a steroid acid or a nutrient. In specificembodiments, the steroid acid or nutrient described herein is a steroidacid or nutrient that enhances the secretion of an enteroendocrinepeptide. In specific embodiments, the steroid acid is an oxidizecholesterol acid. In some embodiments, an enteroendocrine peptidesecretion enhancing agent, bile acid, or bile acid mimic used in anycomposition or method described herein is a compound of Formula VII:

In certain embodiments, each R¹ is independently H, OH, O-lower alkyl(e.g., OCH₃, or OEt). In some embodiments, each R¹ is independently H,OH, lower (e.g., C₁-C₆ or C₁-C₃) alkyl, or lower (e.g., C₁-C₆ or C₁-C₃)heteroalkyl. In certain embodiments, L is a substituted or unsubstitutedalkyl or substituted or unsubstituted heteroalkyl. In some embodiments,R² is H, OH, lower alkyl, or lower heteroalkyl (e.g., OMe). In certainembodiments, R³ is H, OH, O-lower alkyl, lower alkyl, or lowerheteroalkyl (e.g., OMe). In some embodiments, A is COOR⁴, S(O)_(n)R⁴, orOR⁵. In certain embodiments, R⁴ is H, an anion, a pharmaceuticallyacceptable cation (e.g., an alkali metal cation, alkaline earth metalcation, or any other pharmaceutically acceptable cation) substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted aryl, substituted or unsubstitutedheteroaryl, an amino acid, or the like; and n is 1-3. Each R⁵ isindependently selected from lower alkyl and H.

In specific embodiments, L is unsubstituted branched or straight chainalkyl. In more specific embodiments, L is unsubstituted branched orstraight chain lower alkyl. In some embodiments, L is (CR⁵₂)_(m)—CONR⁵—(CR⁵ ₂)_(p). Each m is 1-6 and n is 1-6. In specificembodiments, m is 2 and n is 1. In other specific embodiments, m is 2and n is 2. In certain embodiments, A is COOH or COO—. In someembodiments, A is SO₃H or SO₃—.

In specific embodiments, the compound of Formula VII has a structurerepresented by:

In some embodiments, bile acid mimics include, by way of non-limitingexample,6-methyl-2-oxo-4-thiophen-2-yl-1,2,3,4-tetrahydro-phyrimidine-5-carboxylicacid benzyl ester (or TGR5-binding analogs thereof), oleanolic acid (orTGR5-binding analogs thereof), crataegolic acid,6α-ethyl-23(S)-methylcholic acid (S-EMCA, INT-777),(3R)-3-Hydroxy-3-(2-propen-1-yl)-lup-20(29)-en-28-oic acid hydrate(RG-239), or the like.

In some embodiments, a bile acid mimic is

In certain embodiments, enteroendocrine peptide secretion enhancingagents used in the methods and compositions described herein enhance thesecretion of an enteroendocrine peptide secreted by L-cells (e.g.,GLP-1, GLP-2, PYY, and the like). FIG. 11 (FIGS. 11A and 11B)illustrates the response of enteroendocrine peptides to administrationof bile salts.

In some embodiments, the enteroendocrine peptide secretion enhancingagent is a steroid acid, such as a bile acid/salt, a bile acid/saltmimic, a modified bile acid/salt, or a combination thereof. The bileacids or salts thereof used in the methods and compositions describedherein include, by way of non-limiting example, cholic acid, deoxycholicacid, glycocholic acid, glycodeoxycholic acid, taurocholic acid,taurodihydrofusidate, taurodeoxycholic acid, cholate, glycocholate,deoxycholate, taurocholate, taurodeoxycholate, chenodeoxycholic acid,ursodeoxycholic acid and combinations thereof. In certain embodiments,bile salts used in the methods and compositions described herein arepharmaceutically acceptable salts including, by way of non-limitingexample, the sodium and potassium salts thereof. In specificembodiments, the enteroendocrine peptide secretion enhancing agent is apharmaceutically acceptable bile acid salt including, by way ofnon-limiting example, sodium glycocholate, sodium taurocholate andcombinations thereof. In some embodiments, more than one bile acidand/or salt is used in a methods and/or composition described herein. Incertain embodiments, the bile acid/salt used herein has a low orrelatively low solubility in water.

Although bile acids facilitate digestion and absorption of lipids in thesmall intestine, they are generally used in pharmaceutical formulationsas excipients. As excipients, bile acids find uses as surfactants and/oras agents that enhance the transfer of active components across mucosalmembranes, for systemic delivery of a pharmaceutically active compound.In certain embodiments of the methods and pharmaceutical compositionsdescribed herein, however, a bile acid, a bile acid mimic and/or amodified bile acid is the active agent used to enhance secretion ofenteroendocrine peptides.

In certain specific embodiments, the enteroendocrine peptide secretionenhancing agents used in the methods and compositions described hereinare modified bile acids/salts. In certain embodiments, thebile-acid/salt is modified in such a way so as to inhibit absorption ofthe bile acid/salt across the rectal or colonic mucosa.

In certain embodiments, the enteroendocrine peptide secretion enhancingagents described herein are a glucagon-like peptide secretion enhancingagent. In a specific embodiment, the glucugen-like peptide secretionenhancing agent is a bile acid, a bile acid mimic or a modified bileacid. In some embodiments, the glucagon-like peptide secretion enhancingagents are selected from, by way of non-limiting example, glucagon-likepeptide-1 (GLP-1) secretion enhancing agents or glucagon-like peptide-2(GLP-2) secretion enhancing agents. In some embodiments, theglucagon-like peptide secretion enhancing agents enhance both GLP-1 andGLP-2. In a specific embodiment, the GLP-1 and/or GLP-2 secretionenhancing agent is selected from bile acids, bile acid mimics ormodified bile acids.

In certain embodiments, the enteroendocrine peptide secretion enhancingagent described herein is a pancreatic polypeptide-fold peptidesecretion enhancing agent. In more specific embodiments, the pancreaticpolypeptide-fold peptide secretion enhancing agent is selected from, byway of non-limiting example, peptide YY (PYY) secretion enhancingagents. In specific embodiments, the pancreatic polypeptide-fold peptidesecretion enhancing agent or the PYY secretion enhancing agent isselected from a bile acid, a bile acid mimic, a modified bile acid or afatty acid or salt thereof (e.g., a short chain fatty acid).

In some embodiments, the enteroendocrine peptide secretion enhancingagent is selected from, by way of non-limiting example, carbohydrates,glucose, fats, and proteins. In certain embodiments, the enteroendocrinepeptide secretion enhancing agent is selected from fatty acids,including long chain fatty acids and short chain fatty acids. Shortchain fatty acids and salts include, by way of non-limiting example,propionic acid, butyric acid, propionate, and butyrate.

In some embodiments, the enteroendocrine peptide secretion enhancingagent is selected from, by way of non-limiting example, carbohydrates,glucose, fat, protein, protein hydrolysate, amino acids, nutrients,intestinal peptides, peripheral hormones that participate in energyhomeostasis, such as the adipocyte hormone leptin, bile acids/salts,insulin, gastrin-releasing peptide (GRP), gut peptides, gastric acid,CCK, insulin-like growth factor 1, bombesin, calcitonin-gene-relatedpeptide and combinations thereof that enhance the secretion ofenteroendocrine peptides.

In certain embodiments, the inhibitors of degradation of L-cellenteroendocrine peptide products include DPP-IV inhibitors, TGR5modulators (e.g., TGR5 agonists), or combinations thereof. In certaininstances, the administration of a DPP-IV inhibitor in combination withany of the compounds disclosed herein reduces or inhibits degradation ofGLP-1 or GLP-2. In certain instances, administration of a TGR5 agonistin combination with any of the compounds disclosed herein enhances thesecretion of enteroendocrine peptide products from L-cells. In someinstances, the enteroendocrine peptide enhancing agent agonizes orpartially agonizes bile acid receptors (e.g., TGR5 receptors orFarnesoid-X receptors) on in the gastrointestinal tract.

DPP-IV inhibitors include(2S)-1-{2-[(3-hydroxy-1-adamantyl)amino]acetyl}pyrrolidine-2-carbonitrile(vildagliptin),(3R)-3-amino-1-[9-(trifluoromethyl)-1,4,7,8-tetrazabicyclo[4.3.0]nona-6,8-dien-4-yl]-4-(2,4,5-trifluorophenyl)butan-1-one(sitagliptin),(1S,3S,5S)-2-[(2S)-2-amino-2-(3-hydroxy-1-adamantyl)acetyl]-2-azabicyclo[3.1.0]hexane-3-carbonitrile(saxagliptin), and2-({6-[(3R)-3-aminopiperidin-1-yl]-3-methyl-2,4-dioxo-3,4-dihydropyrimidin-(2H)-yl}methyl)benzonitrile(alogliptin). TGR5 modulators (e.g., agonists) include the compoundsdisclosed in, e.g, WO2008/091540, WO 2008067219 and US Appl. No.2008/0221161, the TGR5 modulators (e.g., agonists) of which are herebyincorporated herein by reference.

In some embodiments, the enteroendocrine peptide secretion enhancingagents used in the methods and compositions described herein may or maynot be substrates for bile acid scavenger systems. In some embodiments,the enteroendocrine peptide secretion enhancing agents may not formmicelles and/or assist in fat absorption. In certain embodiments, theenteroendocrine peptide secretion enhancing agents may or may notenhance permeability and/or promote inflammation. In certainembodiments, the enteroendocrine peptide secretion enhancing agent maynot irritate the bowel or promote diarrhea. In some embodiments, theenteroendocrine peptide secretion enhancing agent is selected from, byway of non-limiting example, toll or toll-like receptor ligands.

FIG. 12 illustrates the affect on food intake of pump infusion of TCA inconcentrations of 150 mM and 500 mM. FIG. 13 illustrates the affect onbody weight of pump infusion of TCA in concentrations of 150 mM and 500mM.

FXR Agonists

In some embodiments, FXR agonist is GW4064, GW9662, 1NT-747, T0901317,WAY-362450, fexaramine, a cholic acid, a deoxycholic acid, a glycocholicacid, a glycodeoxycholic acid, a taurocholic acid, ataurodihydrofusidate, a taurodeoxycholic acid, a cholate, aglycocholate, a deoxycholate, a taurocholate, a taurodeoxycholate, achenodeoxycholic acid.

Absorption Inhibitors

In certain embodiments, the compositions described herein are and themethods described herein include administering a composition that isformulated for the non-systemic delivery of enteroendocrine peptidesecretion enhancing agents to the rectum and/or colon (sigmoid,transverse, and/or ascending colon). As previously discussed,enteroendocrine peptide secretion enhancing agents include, by way ofnon-limiting example, bile acids, bile salts, bile acid mimics, bilesalt mimics, modified bile acids, modified bile salts and combinationsthereof. In certain embodiments, the composition described herein asbeing formulated for the non-systemic delivery of enteroendocrinepeptide secretion enhancing agents further includes an absorptioninhibitor. As used herein, an absorption inhibitor includes an agent orgroup of agents that inhibit absorption of the enteroendocrine peptidesecretion enhancing agent across the rectal or colonic mucosa. Inspecific embodiments, the absorption inhibitor is an absorptioninhibitor that inhibits the absorption of the specific enteroendocrinepeptide secretion enhancing agent with which it is combined.

Suitable bile acid absorption inhibitors (also described herein asabsorption inhibiting agents) include, by way of non-limiting example,anionic exchange matrices, polyamines, quaternary amine containingpolymers, quaternary ammonium salts, polyallylamine polymers andcopolymers, colesevelam, colesevelam hydrochloride, CholestaGel(N,N,N-trimethyl-6-(2-propenylamino)-1-hexanaminium chloride polymerwith (chloromethyl)oxirane, 2-propen-1-amine andN-2-propenyl-1-decanamine hydrochloride), cyclodextrins, chitosan,chitosan derivatives, carbohydrates which bind bile acids, lipids whichbind bile acids, proteins and proteinaceous materials which bind bileacids, and antibodies and albumins which bind bile acids. Suitablecyclodextrins include those that bind bile acids such as, by way ofnon-limiting example, β-cyclodextrin and hydroxypropyl-β-cyclodextrin.Suitable proteins, include those that bind bile acids such as, by way ofnon-limiting example, bovine serum albumin, egg albumin, casein, α-acidglycoprotein, gelatin, soy proteins, peanut proteins, almond proteins,and wheat vegetable proteins.

In certain embodiments the absorption inhibitor is cholestyramine. Inspecific embodiments, cholestyramine is combined with a bile acid.Cholestyramine, an ion exchange resin, is a styrene polymer containingquaternary ammonium groups crosslinked by divinylbenzene. In otherembodiments, the absorption inhibitor is colestipol. In specificembodiments, colestipol is combined with a bile acid. Colestipol, an ionexchange resin, is a copolymer of diethylenetriamine and1-chloro-2,3-epoxypropane.

In certain embodiments of the compositions and methods described hereinthe enteroendocrine peptide secretion enhancing agent is linked to anabsorption inhibitor, while in other embodiments the enteroendocrinepeptide secretion enhancing agent and the absorption inhibitor areseparate molecular entities. In specific embodiments the bile acid, bileacid mimic or the modified bile acid is linked to a bile acid adsorptioninhibitor described herein.

Cholesterol Absorption Inhibitors

In certain embodiments, a composition described herein optionallyincludes at least one cholesterol absorption inhibitor. Suitablecholesterol absorption inhibitors include, by way of non-limitingexample, ezetimibe (SCH 58235), ezetimibe analogs, ACT inhibitors,stigmastanyl phosphorylcholine, stigmastanyl phosphorylcholineanalogues, □-lactam cholesterol absorption inhibitors, sulfatepolysaccharides, neomycin, plant sponins, plant sterols, phytostanolpreparation FM-VP4, Sitostanol, □-sitosterol,acyl-CoA:cholesterol-O-acyltransferase (ACAT) inhibitors, Avasimibe,Implitapide, steroidal glycosides and the like. Suitable enzetimibeanalogs include, by way of non-limiting example, SCH 48461, SCH 58053and the like. Suitable ACT inhibitors include, by way of non-limitingexample, trimethoxy fatty acid anilides such as Cl-976,3-[decyldimethylsilyl]-N-[2-(4-methylphenyl)-1-phenylethyl]-propanamide,melinamide and the like. □-lactam cholesterol absorption inhibitorsinclude, by way of non-limiting example,(3R-4S)-1,4-bis-(4-methoxyphenyl)-3-(3-phenylpropyl)-2-azetidinone andthe like.

Enteroendocrine Peptides

In certain embodiments, the compositions described herein optionallyinclude at least one enteroendocrine peptide. Suitable enteroendocrinepeptides include, by way of non-limiting example, glucagon-like peptidesGLP-1 and/or GLP-2, or pancreatic polypeptide-fold peptides pancreaticpolypeptide (PP), neuropeptide Y (NPY) and/or peptide YY (PYY).

Peptidase Inhibitors

In some embodiments, the compositions described herein optionallyinclude at least one peptidase inhibitor. Such peptidase inhibitorsinclude, but are not limited to, dipeptidyl peptidase-4 inhibitors(DPP-4), neutral endopeptidase inhibitors, and converting enzymeinhibitors. Suitable dipeptidyl peptidase-4 inhibitors (DPP-4) include,by way of non-limiting example, Vildaglipti,2S)-1-{2-[(3-hydroxy-1-adamantyl)amino]acetyl}pyrrolidine-2-carbonitrile,Sitagliptin,(3R)-3-amino-1-[9-(trifluoromethyl)-1,4,7,8-tetrazabicyclo[4.3.0]nona-6,8-dien-4-yl]-4-(2,4,5-trifluorophenyl)butan-1-one,Saxagliptin, and(1S,3S,5S)-2-[(2S)-2-amino-2-(3-hydroxy-1-adamantyl)acetyl]-2-azabicyclo[3.1.0]hexane-3-carbonitrile.Such neutral endopeptidase inhibitors include, but are not limited to,Candoxatrilat and Ecadotril.

Spreading Agents/Wetting Agents

In certain embodiments, the composition described herein optionallycomprises a spreading agent. In some embodiments, a spreading agent isutilized to improve spreading of the composition in the colon and/orrectum. Suitable spreading agents include, by way of non-limitingexample, hydroxyethylcellulose, hydroxypropymethyl cellulose,polyethylene glycol, colloidal silicon dioxide, propylene glycol,cyclodextrins, microcrystalline cellulose, polyvinylpyrrolidone,polyoxyethylated glycerides, polycarbophil, di-n-octyl ethers,Cetiol™OE, fatty alcohol polyalkylene glycol ethers, Aethoxal™B),2-ethylhexyl palmitate, Cegesoft™C 24), and isopropyl fatty acid esters.

In some embodiments, the compositions described herein optionallycomprise a wetting agent. In some embodiments, a wetting agent isutilized to improve wettability of the composition in the colon andrectum. Suitable wetting agents include, by way of non-limiting example,surfactants. In some embodiments, surfactants are selected from, by wayof non-limiting example, polysorbate (e.g., 20 or 80), stearylhetanoate, caprylic/capric fatty acid esters of saturated fatty alcoholsof chain length C₁₂-C₁₈, isostearyl diglycerol isostearic acid, sodiumdodecyl sulphate, isopropyl myristate, isopropyl palmitate, andisopropyl myristate/isopropyl stearate/isopropyl palmitate mixture.

Methods

Provided herein, in certain embodiments, are methods for treatingobesity, diabetes, or an inflammatory intestinal condition comprisingadministration of a therapeutically effective amount of an ASBTI and/oran enteroendocrine peptide enhancing agent and/or a FXR agonist to anindividual in need thereof. Provided herein, in certain embodiments, aremethods for treating obesity, diabetes, or an inflammatory intestinalcondition comprising contacting the distal gastrointestinal tract,including the distal ileum and/or the colon and/or the rectum, of anindividual in need thereof with an ASBTI and/or an enteroendocrinepeptide enhancing agent and/or a FXR agonist. Also provided herein aremethods for reducing intraenterocyte bile acids, reducing necrosisand/or damage to ileal architecture, or reducing blood or plasma glucoselevels of an individual comprising administration of a therapeuticallyeffective amount of an ASBTI and/or an enteroendocrine peptide enhancingagent and/or a FXR agonist to an individual in need thereof. Providedherein are methods for stimulating L-cells in the distalgastrointestinal tract, including L-cells in the distal ileum and/orcolon and/or rectum, of an individual comprising administration of atherapeutically effective amount of an ASBTI and/or an enteroendocrinepeptide enhancing agent and/or a FXR agonist to an individual in needthereof. Provided herein are methods for enhancing enteroendocrinepeptide secretion from L-cells in the distal gastrointestinal tract,including L-cells in the distal ileum and/or colon and/or rectum, of anindividual comprising administration of a therapeutically effectiveamount of an ASBTI and/or an enteroendocrine peptide enhancing agentand/or a FXR agonist to an individual in need thereof. Provided hereinare methods for increasing the concentration of bile acids and saltsthereof in the vicinity of L-cells lining the distal gastrointestinaltract, including L-cells in the distal ileum, and/or the colon and/orthe rectum of an individual, comprising administration of atherapeutically effective amount of an ASBTI and/or an enteroendocrinepeptide enhancing agent and/or a FXR agonist to an individual in needthereof. In some of the aforementioned embodiments, the ASBTI iscontacted with the distal ileum of the individual in need thereof. Insome of the aforementioned embodiments, the ASBTI is not absorbedsystemically. In some other embodiments, the ASBTI is absorbedsystemically.

In some embodiments of the methods provided herein, inhibition of bileacid transporters and/or bile acid recycling increases the concentrationof bile acids in the vicinity of L-cells to concentrations that arehigher than physiological levels of bile acids in individuals that havenot been treated with an ASBTI. In some embodiments of the methodsdescribed herein, an increase in concentration of bile acids in thevicinity of L-cell increases the secretion of enteroendocrine peptides,including GLP-1, GLP-2, PYY and/or oxyntomodulin from L-cells. In someinstances a higher concentration of GLP-1 and/or GLP-2 and/or PYY and/oroxynotmodulin in the blood and/or plasma of an individual, increasesinsulin sensitivity of the individual, reduces intraenterocyte bileacids, reduces necrosis and/or damage to ileal architecture, and/orslows down gastric emptying and/or induces a feeling of satiety therebyreducing food intake and/or inducing weight loss.

Provided herein are methods for reducing necrosis and/or damage to ilealarchitecture or cells comprising administration of a therapeuticallyeffective amount of an ASBTI and/or an enteroendocrine peptide enhancingagent and/or a FXR agonist to an individual in need thereof. In certainembodiments, provided herein are methods for reducing intraenterocytebile acids comprising administration of a therapeutically effectiveamount of an ASBTI and/or an enteroendocrine peptide enhancing agentand/or a FXR agonist to an individual in need thereof. In certainembodiments, provided herein are methods for reducing food intake in anindividual comprising administration of a therapeutically effectiveamount of an ASBTI to an individual in need thereof. In certainembodiments, provided herein is a method for reducing food intake in anindividual in need thereof comprising contacting the distalgastrointestinal tract, including the distal ileum and/or colon and/orrectum, of the individual with a therapeutically effective amount of anASBTI. Provided herein are methods for reducing weight of an individualcomprising administration of a therapeutically effective amount of anASBTI to an individual in need thereof. In certain embodiments, providedherein is a method of reducing weight of an individual in need thereofcomprising contacting the distal gastrointestinal tract, including thedistal ileum and/or colon and/or rectum, of the individual with atherapeutically effective amount of an ASBT inhibitor. In someembodiments, the method provides for inhibition of bile salt recyclingupon administration of any of the compounds described herein to anindividual. In some embodiments, an ASBTI described herein issystemically absorbed upon administration. In some embodiments, an ASBTIdescribed herein is not absorbed systemically. In some embodiments, anASBTI described herein is administered to the individual orally,enterically or rectally. In some embodiments, an ASBTI described hereinis delivered and/or released in the distal ileum of an individual. Insome embodiments, an ASBTI described herein increases the concentrationof bile acids in the distal ileum, the colon and/or the rectum therebyincreasing secretion of enteroendocrine peptide products from L-cells inthe gastrointestinal tract. In certain instances administration of atherapeutically effective amount of an ASBTI described herein to anindividual in need thereof increases the secretion of enteroendocrinepeptide products (e.g., GLP-1, GLP-2, PYY, oxytonmodulin or the like)from L-cells that line the gastrointestinal tract. In some embodiments,elevated levels of GLP-1 slow down gastric emptying, and/or inhibit orreduce meal-stimulated gastric secretion, thereby reducing food intakein the individual. In some embodiments, an ASBTI described herein isadministered in combination with a DPP-IV inhibitor. In some instances,inhibition of DPP-IV reduces the degradation of enteroendocrine peptideproducts (e.g. GLP-1) thereby prolonging the delay in gastric emptyingand thereby reducing food intake. In some of such embodiments, an ASBTIdescribed herein is administered to a non-diabetic individual forreducing food intake in the non-diabetic individual. In some of suchembodiments, an ASBTI described herein is administered to a diabeticindividual for reducing food intake in the diabetic individual. In someembodiments, the methods described herein are methods for reducing foodintake in obese or morbidly overweight individuals. In some embodiments,a reduction in food intake reduces the weight of an individual (e.g., anobese or morbidly overweight individual)

Provided herein are methods for inducing satiety in an individualcomprising administration of a therapeutically effective amount of anASBTI to an individual in need thereof. In certain embodiments, providedherein is a method for inducing satiety in an individual in need thereofcomprising contacting the distal gastrointestinal tract, includingdistal ileum and/or the colon and/or the rectum, of the individual witha therapeutically effective amount of an ASBT inhibitor. In someembodiments, the method provides for inhibition of bile salt recyclingupon administration of any of the compounds described herein to anindividual. In some embodiments, an ASBTI described herein issystemically absorbed upon administration. In some embodiments, an ASBTIdescribed herein (e.g., a compound of Formula I) is not absorbedsystemically. In some embodiments, an ASBTI described herein isadministered to the individual orally, enterically or rectally. In someembodiments, an ASBTI described herein is delivered and/or released inthe distal ileum of an individual. In some embodiments, an ASBTIdescribed herein increases the concentration of bile acids in the distalileum, the colon and/or the rectum and induces secretion ofenteroendocrine peptide products from L-cells of the distalgastrointestinal tract. In certain instances administration of atherapeutically effective amount of an ASBTI described herein to anindividual in need thereof increases the secretion of enteroendocrinepeptide products (e.g., GLP-1, GLP-2, PYY, oxytonmodulin or the like)from L-cells that line the gastrointestinal tract. In some embodiments,elevated levels of GLP-1 slow down gastric emptying, and induce afeeling of fullness in an individual. In some embodiments, an ASBTIdescribed herein is administered in combination with a DPP-IV inhibitor.In some instances, inhibition of DPP-IV reduces the degradation ofenteroendocrine peptide products (e.g. GLP-1) thereby prolonging thedelay in gastric emptying and sustaining the feeling of satiety and/orfullness. In some of such embodiments, an ASBTI described herein isadministered to a non-diabetic individual. In some of such embodiments,an ASBTI described herein is administered to a diabetic individual. Insome of such embodiments, an ASBTI described herein is administered toan obese or morbidly overweight individual.

Provided herein are methods for preventing or treating metabolicdisorders in an individual comprising administration of atherapeutically effective amount of an ASBTI and/or an enteroendocrinepeptide enhancing agent and/or a FXR agonist to an individual in needthereof. In certain embodiments, the metabolic disorders include but notlimited to necrotizing enterocolitis, gastritis, ulcerative colitis,inflammatory bowel disease, irritable bowel syndrome, gastroenteritis,radiation induced enteritis, chemotherapy induced enteritis,gastro-esophageal reflux disease (GERD), peptic ulcer, non-ulcerdyspepsia (NUD), celiac disease, intestinal celiac disease,gastrointestinal complications following bariatric surgery, gastriccarcinogenesis, or gastric carcinogenesis following gastric or bowelresection. In certain embodiments, provided herein is a method fortreating metabolic disorders in an individual in need thereof comprisingcontacting the distal gastrointestinal tract, including distal ileumand/or the colon and/or the rectum, of the individual with atherapeutically effective amount of an ASBTI and/or an enteroendocrinepeptide enhancing agent and/or a FXR agonist. In some embodiments, themethod provides for inhibition of bile salt recycling uponadministration of any of the compounds described herein to anindividual. In some embodiments, an ASBTI described herein issystemically absorbed upon administration. In some embodiments, an ASBTIdescribed herein (e.g., a compound of Formula I) is not absorbedsystemically. In some embodiments, an ASBTI described herein isadministered to the individual orally, enterically or rectally. In someembodiments, an ASBTI and/or an enteroendocrine peptide enhancing agentand/or a FXR agonist described herein is delivered and/or released inthe distal ileum of an individual. In some embodiments, an ASBTI and/oran enteroendocrine peptide enhancing agent and/or a FXR agonistdescribed herein increases the concentration of bile acids in the distalileum, the colon and/or the rectum and induces secretion ofenteroendocrine peptide products from the L-cells of thegastrointestinal tract. In certain instances administration of atherapeutically effective amount of an ASBTI described herein to anindividual in need thereof increases the secretion of enteroendocrinepeptide products (e.g., GLP-1, GLP-2, PYY, oxytonmodulin or the like)from L-cells that line the distal gastrointestinal tract. In someembodiments, elevated levels of GLP-1 reduce glucose levels in blood. Insome instances, elevated levels of GLP-1 increase insulin sensitivity ina hyperglycemic individual. In some embodiments, an ASBTI and/or anenteroendocrine peptide enhancing agent and/or a FXR agonist describedherein is administered in combination with a DPP-IV inhibitor. In someinstances, inhibition of DPP-IV reduces the degradation ofenteroendocrine peptide products (e.g. GLP-1) thereby prolonging theeffect of GLP-1 in reducing blood glucose levels. In some of suchembodiments, an ASBTI described herein is administered to a non-diabeticindividual. In some of such embodiments, an ASBTI described herein isadministered to a diabetic individual. In some of such embodiments, anASBTI described herein is administered to an obese or morbidlyoverweight individual.

Provided herein are methods for preventing or treating radiation inducedenteritis in an individual comprising administration of atherapeutically effective amount of an ASBTI and/or an enteroendocrinepeptide enhancing agent and/or a FXR agonist to an individual in needthereof. In certain embodiments, provided herein is a method fortreating radiation enteritis in an individual in need thereof comprisingcontacting the distal gastrointestinal tract, including distal ileumand/or the colon and/or the rectum, of the individual with atherapeutically effective amount of an ASBTI and/or an enteroendocrinepeptide enhancing agent and/or a FXR agonist. In some embodiments, themethod provides for inhibition of bile salt recycling uponadministration of any of the compounds described herein to anindividual. In some embodiments, an ASBTI described herein issystemically absorbed upon administration. In some embodiments, an ASBTIdescribed herein (e.g., a compound of Formula I) is not absorbedsystemically. In some embodiments, an ASBTI and/or an enteroendocrinepeptide enhancing agent and/or a FXR agonist described herein isadministered to the individual orally, enterically or rectally. In someembodiments, an ASBTI described herein is delivered and/or released inthe distal ileum of an individual. In some embodiments, an ASBTI and/oran enteroendocrine peptide enhancing agent and/or a FXR agonistdescribed herein increases the concentration of bile acids in the distalileum, the colon and/or the rectum and induces secretion ofenteroendocrine peptide products from the L-cells of thegastrointestinal tract. In certain instances administration of atherapeutically effective amount of an ASBTI described herein to anindividual in need thereof increases the secretion of enteroendocrinepeptide products (e.g., GLP-1, GLP-2, PYY, oxytonmodulin or the like)from L-cells that line the distal gastrointestinal tract. In someembodiments, elevated levels of GLP-2 has a regenerative effect onradiation-induced injuries of the gastrointestinal tract (e.g., afterradiation therapy for treatment of cancer, or accidental exposure toradiation). In some embodiments, prophylactic administration of an ASBTIand/or an enteroendocrine peptide enhancing agent and/or a FXR agonistreduces or prevents intestinal inflammation (gastrointestinalenteritis), for example in a cancer patient undergoing radiationtherapy. In some embodiments, an ASBTI described herein is administeredin combination with a DPP-IV inhibitor. In some instances, inhibition ofDPP-IV reduces the degradation of enteroendocrine peptide products (e.g.GLP-2) thereby prolonging the effect of GLP-2 in regeneration and/orhealing of gastronintestinal tissue.

Provided herein are methods for preventing or treating pancreatic andother cancers in an individual comprising administration of atherapeutically effective amount of an ASBTI and/or an enteroendocrinepeptide enhancing agent and/or a FXR agonist to an individual in needthereof. Other cancers include but are not limited to colon, breast,bowel and lung. In certain embodiments, provided herein is a method forpreventing or treating pancreatic and other cancers cancer in anindividual in need thereof comprising contacting the distalgastrointestinal tract, including distal ileum and/or the colon and/orthe rectum, of the individual with a therapeutically effective amount ofan ASBTI. In some embodiments, the method provides for inhibition ofbile salt recycling upon administration of any of the compoundsdescribed herein to an individual. In some embodiments, an ASBTIdescribed herein is systemically absorbed upon administration. In someembodiments, an ASBTI described herein (e.g., a compound of Formula I)is not absorbed systemically. In some embodiments, an ASBTI and/or anenteroendocrine peptide enhancing agent and/or a FXR agonist describedherein is administered to the individual orally, enterically orrectally. In some embodiments, an ASBTI described herein is deliveredand/or released in the distal ileum of an individual. In someembodiments, an ASBTI and/or an enteroendocrine peptide enhancing agentand/or a FXR agonist described herein increases the concentration ofbile acids in the distal ileum, the colon and/or the rectum and inducessecretion of enteroendocrine peptide products from the L-cells of thegastrointestinal tract. In certain instances administration of atherapeutically effective amount of an ASBTI described herein to anindividual in need thereof increases the secretion of enteroendocrinepeptide products (e.g., GLP-1, GLP-2, PYY, oxytonmodulin or the like)from L-cells that line the distal gastrointestinal tract. In someembodiments, elevated levels of enteroendocrine peptide products have aprotective effect against pancreatic and other cancers. In someembodiments, prophylactic administration of an ASBTI reduces or preventspancreatic and other cancers. In some embodiments, an ASBTI and/or anenteroendocrine peptide enhancing agent and/or a FXR agonist describedherein is administered in combination with a DPP-IV inhibitor. In someinstances, inhibition of DPP-IV reduces the degradation ofenteroendocrine peptide products (e.g., GLP-1) thereby prolonging theeffect of GLP-1 in prevention of pancreatic and other cancers. In someof such embodiments, an ASBTI described herein is administered to anindividual with pancreatic and other cancers. In some of suchembodiments, an ASBTI described herein is administered to an individualat risk of pancreatic and other cancers. In some of such embodiments, anASBTI described herein is administered to an individual at risk ofpancreatic and other cancers and having other metabolic disorders (e.g.,diabetes).

In some embodiments, administration of an ASBT inhibitor and/or anenteroendocrine peptide enhancing agent and/or a FXR agonist describedherein allows for treatment of a metabolic disorder without the sideeffects associated with conventional therapies (e.g., biguanides such asmetformin, DDPIV inhibitors or the like) for metabolic disorders. Insome embodiments, the administration of an ASBT inhibitor describedherein reduces the incidence of gastrointestinal distress and/or lacticacidosis that is associated with biguanide therapy and/or treatment witha DPP-IV inhibitor. In some embodiments, the administration of an ASBTinhibitor described herein avoids weight gain that is associated withbiguanide therapy and/or treatment with a DPP-IV inhibitor and/or anenteroendocrine peptide enhancing agent and/or a FXR agonist. Metabolicdisorders that are amenable to treatment with compounds described hereininclude diabetes, insulin resistance, and metabolic syndrome.

In some embodiments, provided herein is a method of decreasing the doseof a biguanide or a DPP-IV inhibitor to treat a metabolic disorder in anindividual in need thereof by replacing treatment with a biguanide(e.g., metformin) or a DPP-IV inhibitor with a treatment comprisingadministering a therapeutically effective amount of an ASBT inhibitordescribed herein to the individual. In some embodiments, provided hereinis a method for treating a metabolic disease or disorder comprisingadministering to an individual in need thereof a therapeuticallyeffective amount of an ASBT inhibitor described herein in combinationwith a reduced dose of a biguanide (e.g., metformin) or a DPP-IVinhibitor (e.g., sitagliptin) or an enteroendocrine peptide enhancingagent or a FXR agonist.

In some embodiments of any of the methods described herein,administration of an ASBT inhibitor and/or an enteroendocrine peptideenhancing agent and/or a FXR agonist described herein increases thelevel of GLP-1 in the blood and/or plasma of an individual by from about1.1 times to about 30 times compared to the level of GLP-1 in the bloodand/or plasma of the individual prior to administration of the ASBTI. Insome embodiments of any of the methods described herein, administrationof an ASBT inhibitor described herein increases the level of GLP-1 inthe blood and/or plasma of an individual by from about 1.1 times toabout 20 times compared to the level of GLP-1 in the blood and/or plasmaof the individual prior to administration of the ASBTI. In someembodiments of any of the methods described herein, administration of anASBT inhibitor and/or an enteroendocrine peptide enhancing agent and/ora FXR agonist described herein increases the level of GLP-1 in the bloodand/or plasma of an individual by from about 1.5 times to about 10 timescompared to the level of GLP-1 in the blood and/or plasma of theindividual prior to administration of the ASBTI and/or anenteroendocrine peptide enhancing agent and/or a FXR agonist. In someembodiments of any of the methods described herein, administration of anASBT inhibitor and/or an enteroendocrine peptide enhancing agent and/ora FXR agonist described herein increases the level of GLP-1 in the bloodand/or plasma of an individual by from about 2 times to about 8 timescompared to the level of GLP-1 in the blood and/or plasma of theindividual prior to administration of the ASBTI and/or anenteroendocrine peptide enhancing agent and/or a FXR agonist. In someembodiments of any of the methods described herein, administration of anASBT inhibitor and/or an enteroendocrine peptide enhancing agent and/ora FXR agonist described herein increases the level of GLP-1 in the bloodand/or plasma of an individual by from about 2 times to about 6 timescompared to the level of GLP-1 in the blood and/or plasma of theindividual prior to administration of the ASBTI and/or anenteroendocrine peptide enhancing agent and/or a FXR agonist. In someinstances, an increase in GLP-1 level of from about 2 times to about 3times following the administration of an ASBT inhibitor and/or anenteroendocrine peptide enhancing agent and/or a FXR agonist describedherein compared to the level of GLP-1 in the blood and/or plasma of theindividual prior to administration of the ASBTI and/or anenteroendocrine peptide enhancing agent and/or a FXR agonist isassociated with an anti-diabetic effect. In some instances, an increasein GLP-1 level of from about 3 times to about 8 times following theadministration of an ASBT inhibitor and/or an enteroendocrine peptideenhancing agent and/or a FXR agonist described herein compared to thelevel of GLP-1 in the blood and/or plasma of the individual prior toadministration of the ASBTI and/or an enteroendocrine peptide enhancingagent and/or a FXR agonist is associated with reduction in food intakeand/or induction of satiety and/or weight loss.

In certain embodiments of any of the methods described herein,administration of an ASBTI and/or an enteroendocrine peptide enhancingagent and/or a FXR agonist reduces blood and/or plasma sugar levels byat least 20%, at least 30%, at least 40%, at least 50% at least 60%, atleast 70% or at least 80% compared to blood and/or plasma sugar levelsprior to administration of the ASBTI and/or an enteroendocrine peptideenhancing agent and/or a FXR agonist. In some embodiments of any of themethods described herein, administration of an ASBTI and/or anenteroendocrine peptide enhancing agent and/or a FXR agonist reducesblood and/or plasma sugar levels by at least 20% compared to bloodand/or plasma sugar levels prior to administration of the ASBTI and/oran enteroendocrine peptide enhancing agent and/or a FXR agonist. In someembodiments of any of the methods described herein, administration of anASBTI and/or an enteroendocrine peptide enhancing agent and/or a FXRagonist reduces blood and/or plasma sugar levels by at least 30%compared to blood and/or plasma sugar levels prior to administration ofthe ASBTI and/or an enteroendocrine peptide enhancing agent and/or a FXRagonist. In some embodiments of any of the methods described herein,administration of an ASBTI and/or an enteroendocrine peptide enhancingagent and/or a FXR agonist reduces blood and/or plasma sugar levels byat least 40% compared to blood and/or plasma sugar levels prior toadministration of the ASBTI and/or an enteroendocrine peptide enhancingagent and/or a FXR agonist.

In some embodiments of any of the methods described herein,administration of an ASBTI reduces blood and/or plasma sugar levels fora longer period of time (e.g., at least 24 hours) compared to reductionin blood and/or plasma sugar levels upon administration of metformin ora DPP-IV inhibitor. In some embodiments of any of the methods describedherein, administration of a single dose of an ASBTI sustains reducedblood and/or plasma sugar levels for at least 6 hours, at least 12hours, at least 14 hours, at least 16 hours, at least 18 hours, at least20 hours, at least 24 hours, at least 30 hours, at least 36 hours or atleast 48 hours compared to reduction in blood and/or plasma sugar levelsupon administration of a single dose of metformin or a DPP-IV inhibitor.

In some embodiments of any of the methods described herein,administration of an ASBTI and/or an enteroendocrine peptide enhancingagent and/or a FXR agonist results in higher levels of GLP-1 in bloodand/or plasma of an individual compared to levels of GLP-1 in bloodand/or plasma of a normal individual. In some embodiments of any of themethods described herein, administration of an ASBTI and/or anenteroendocrine peptide enhancing agent and/or a FXR agonist results inhigher levels of GLP-1 in blood and/or plasma of an individual comparedto levels of GLP-1 in blood and/or plasma of an individual that has beenadministered a DPP-IV inhibitor.

Also provided herein is a method for treating conditions that areameliorated by increased secretion of L-cell enteroendocrine peptidescomprising contacting the distal gastrointestinal tract, includingdistal ileum and/or the colon and/or the rectum, of an individual inneed thereof with a therapeutically effective amount of any ASBTIcompound and/or an enteroendocrine peptide enhancing agent and/or a FXRagonist described herein. L-cells are highly specialized gutenteroendocrine cells expressed along the gastrointestinal tract. Themajority of L cells are located in the distal gastrointestinal tract,predominantly in the ileum and colon. The L-cells in the entericendocrine system do not secrete their hormone continuously. Instead,they respond to changes in the environment within the lumen of thedigestive tube, including changes in bile acid concentrations in thelumen of the digestive tube. The apical border of L-cells is in contactwith the contents of the gastrointestinal lumen. Enteroendocrinepeptides secreted by L-cells include GLP-1, GLP-2, PYY andoxyntomodulin. In certain instances, the methods described hereinenhance L-cell secretion of one or more metabolic and/or enteroendocrinehormones. In some embodiments, the methods described herein enhanceL-cell secretion of GLP-1, GLP-2, PYY or oxyntomodulin or combinationsthereof. In certain embodiments, enhanced secretion of multipleenteroendocrine hormones (e.g., enhanced secretion of PYY and/or GLP-1and/or GLP-2 and/or oxyntomodulin) is more effective for long termweight reduction compared to enhanced secretion of any singleenteroendocrine hormone. In certain embodiments, enhanced secretion ofmultiple enteroendocrine hormones (e.g., enhanced secretion of PYYand/or GLP-1 and/or GLP-2 and/or oxyntomodulin) is more effective forlong term improvement of peripheral insulin sensitivity compared toenhanced secretion of any single enteroendocrine hormone. Thus,advantageously, administration of a single therapeutic agent (e.g., anyASBTI described herein) has the potential to simultaneously impactmultiple metabolic pathways (as opposed to administration of multipletherapeutic agents each of which impacts a different metabolic pathway).

In certain instances, contacting the distal ileum of an individual withan ASBTI (e.g., any ASBTI described herein) inhibits bile acid reuptakeand increases the concentration of bile acids in the vicinity of L-cellsin the distal ileum and/or colon and/or rectum, thereby reducingintraenterocyte bile acids, enhancing the release of enteroendocrinepeptides, and/or reducing necrosis or damage to ileal architecture.Without being limited to any particular theory, bile acids and/or bilesalts interact with TGR5 receptors on the apical surface of L-cells totrigger the release of one or more enteroendocrine hormones intosystemic circulation and/or the gastrointestinal lumen. Underphysiological conditions, the concentration of enteroendocrine hormonesvaries in the gastrointestinal tract. By way of example, in the absenceof an ASBTI, PYY concentrations in the upper small intestine are about˜5 pmol/g tissue, about ˜80 pmol/g tissue in the distal ileum andascending colon, ˜200 pmol/g tissue in the sigmoid colon, and ˜500pmol/g tissue in the rectum. In some embodiments, the administration ofone or more ASBTIs, according to methods described herein, increasesconcentrations of one or more enteroendocrine peptides in thegastrointestinal lumen and/or systemic circulation compared tophysiological concentrations of the enteroendocrine peptides in theabsence of an ASBTI.

Conditions that are mediated by L-cell enteroendocrine peptides includeobesity, diabetes, congestive heart failure, ventricular dysfunction,toxic hypervolemia, polycystic ovary syndrome, inflamatory boweldisease, impaired bowel integrity, short bowel syndrome, gastritis,peptic ulcer, necrotizing enterocolitis, ulcerative colitis, celiacdisease, intestinal celiac disease, gastroenteritis, radiation inducedenteritis, chemotherapy induced enteritis, non-ulcer dyspepsia (NUD),gastrointestinal complications following bariatric surgery, gastriccarcinogenesis, irritable bowel disease, gastroesophageal reflux disease(GERD), Barrett's esophagus or the like.

Administration of a compound described herein is achieved in anysuitable manner including, by way of non-limiting example, by oral,enteric, parenteral (e.g., intravenous, subcutaneous, intramuscular),intranasal, buccal, topical, rectal, or transdermal administrationroutes. Any compound or composition described herein is administered ina method or formulation appropriate to treat a new born or an infant.Any compound or composition described herein is administered in an oralformulation (e.g., solid or liquid) to treat a new born or an infant.Any compound or composition described herein is administered prior toingestion of food, with food or after ingestion of food.

In certain embodiments, a compound or a composition comprising acompound described herein is administered for prophylactic and/ortherapeutic treatments. In therapeutic applications, the compositionsare administered to an individual already suffering from a disease orcondition, in an amount sufficient to cure or at least partially arrestthe symptoms of the disease or condition. In various instances, amountseffective for this use depend on the severity and course of the diseaseor condition, previous therapy, the individual's health status, weight,and response to the drugs, and the judgment of the treating physician.

In prophylactic applications, compounds or compositions containingcompounds described herein are administered to an individual susceptibleto or otherwise at risk of a particular disease, disorder or condition.In certain embodiments of this use, the precise amounts of compoundadministered depend on the individual's state of health, weight, and thelike. Furthermore, in some instances, when a compound or compositiondescribed herein is administered to an individual, effective amounts forthis use depend on the severity and course of the disease, disorder orcondition, previous therapy, the individual's health status and responseto the drugs, and the judgment of the treating physician.

In certain instances, wherein following administration of a selecteddose of a compound or composition described herein, an individual'scondition does not improve, upon the doctor's discretion theadministration of a compound or composition described herein isoptionally administered chronically, that is, for an extended period oftime, including throughout the duration of the individual's life inorder to ameliorate or otherwise control or limit the symptoms of theindividual's disorder, disease or condition.

In certain embodiments, an effective amount of a given agent variesdepending upon one or more of a number of factors such as the particularcompound, disease or condition and its severity, the identity (e.g.,weight) of the subject or host in need of treatment, and is determinedaccording to the particular circumstances surrounding the case,including, e.g., the specific agent being administered, the route ofadministration, the condition being treated, and the subject or hostbeing treated. In some embodiments, doses administered include those upto the maximum tolerable dose. In some embodiments, doses administeredinclude those up to the maximum tolerable dose by a newborn or aninfant. In certain embodiments, about 0.001-5000 mg per day, from about0.001-1500 mg per day, about 0.001 to about 100 mg/day, about 0.001 toabout 50 mg/day, or about 0.001 to about 30 mg/day, or about 0.001 toabout 10 mg/day of a compound described herein is administered. Invarious embodiments, the desired dose is conveniently presented in asingle dose or in divided doses administered simultaneously (or over ashort period of time) or at appropriate intervals, for example as two,three, four or more sub-doses per day. In various embodiments, a singledose is from about 0.001 mg/kg to about 500 mg/kg. In variousembodiments, a single dose is from about 0.001, 0.01, 0.1, 1, or 10mg/kg to about 10, 50, 100, or 250 mg/kg. In various embodiments, asingle dose of an ASBTI is from about 0.001 mg/kg to about 100 mg/kg. Invarious embodiments, a single dose of an ASBTI is from about 0.001 mg/kgto about 50 mg/kg. In various embodiments, a single dose of an ASBTI isfrom about 0.001 mg/kg to about 10 mg/kg. In various embodiments, asingle dose of an ASBTI is administered every 6 hours, every 12 hours,every 24 hours, every 48 hours, every 72 hours, every 96 hours, every 5days, every 6 days, or once a week. In some embodiments the total singledose of an ASBTI and/or an enteroendocrine peptide enhancing agentand/or a FXR agonist is in the range described above.

In the case wherein the patient's status does improve, upon the doctor'sdiscretion an ASBTI is optionally given continuously; alternatively, thedose of drug being administered is temporarily reduced or temporarilysuspended for a certain length of time (i.e., a “drug holiday”). Thelength of the drug holiday optionally varies between 2 days and 1 year,including by way of example only, 2 days, 3 days, 4 days, 5 days, 6days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250days, 280 days, 300 days, 320 days, 350 days, or 365 days. The dosereduction during a drug holiday includes from 10%-100%, including, byway of example only, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments thetotal single dose of an ASBTI and/or an enteroendocrine peptideenhancing agent and/or a FXR agonist is in the range described above.

Once improvement of the patient's conditions has occurred (e.g., weightloss), a maintenance dose is administered if necessary. Subsequently,the dosage or the frequency of administration, or both, is reduced, as afunction of the symptoms, to a level at which the improved disease,disorder or condition is retained. In some embodiments, patients requireintermittent treatment on a long-term basis upon any recurrence ofsymptoms (e.g., weight gain).

In certain instances, there are a large number of variables in regard toan individual treatment regime, and considerable excursions from theserecommended values are considered within the scope described herein.Dosages described herein are optionally altered depending on a number ofvariables such as, by way of non-limiting example, the activity of thecompound used, the disease or condition to be treated, the mode ofadministration, the requirements of the individual subject, the severityof the disease or condition being treated, and the judgment of thepractitioner.

Toxicity and therapeutic efficacy of such therapeutic regimens areoptionally determined by pharmaceutical procedures in cell cultures orexperimental animals, including, but not limited to, the determinationof the LD₅₀ (the dose lethal to 50% of the population) and the ED₅₀ (thedose therapeutically effective in 50% of the population). The dose ratiobetween the toxic and therapeutic effects is the therapeutic index andit can be expressed as the ratio between LD₅₀ and ED₅₀. Compoundsexhibiting high therapeutic indices are preferred. In certainembodiments, data obtained from cell culture assays and animal studiesare used in formulating a range of dosage for use in human. In specificembodiments, the dosage of compounds described herein lies within arange of circulating concentrations that include the ED₅₀ with minimaltoxicity. The dosage optionally varies within this range depending uponthe dosage form employed and the route of administration utilized.

In some embodiments, the systemic exposure of a therapeuticallyeffective amount of any non-systemic ASBTI described herein (e.g., anASBTI that comprises a group L-K) is reduced when compared to thesystemic exposure of a therapeutically effective amount of anysystemically absorbed ASBTI (e.g. Compounds 100A, 100C). In someembodiments, the AUC of a therapeutically effective amount of anynon-systemic ASBTI described herein (e.g., an ASBTI that comprises agroup L-K) is at least 10%, at least 20%, at least 30%, at least 40%, atleast 50%, at least 60%, at least 70%, at least 80% or at least 90%reduced when compared to the AUC of any systemically absorbed ASBTI(e.g. Compounds 100A, 100C).

In some embodiments, the systemic exposure of a therapeuticallyeffective amount of a compound of Formula I that is not systemicallyabsorbed (e.g., a compound of Formula I that comprises a group L-K) isreduced when compared to the systemic exposure of a therapeuticallyeffective amount of Compound 1 OA. In some embodiments, the AUC of atherapeutically effective amount of a compound of Formula I that is notsystemically absorbed (e.g., a compound of Formula I that comprises agroup L-K) is about 10%, about 20%, about 30%, about 40%, about 50%,about 60%, about 70%, about 80% or about 90% reduced when compared tothe AUC of a therapeutically effective amount of Compound 100A. In someembodiments, the AUC of a therapeutically effective amount of a compoundof Formula I that is not systemically absorbed (e.g., a compound ofFormula I that comprises a group L-K) is about 50% reduced when comparedto the AUC of a therapeutically effective amount of Compound 100A. Inother embodiments, the AUC of a therapeutically effective amount of acompound of Formula I that is not systemically absorbed (e.g., acompound of Formula I that comprises a group L-K) is about 75% reducedwhen compared to the AUC of a therapeutically effective amount ofCompound 100A.

In some embodiments, the systemic exposure of a therapeuticallyeffective amount of a compound of Formula II that is not systemicallyabsorbed (e.g., a compound of Formula II that comprises a group L-K) isreduced when compared to the systemic exposure of a therapeuticallyeffective amount of Compound 100A. In some embodiments, the AUC of atherapeutically effective amount of a compound of Formula II that is notsystemically absorbed (e.g., a compound of Formula II that comprises agroup L-K) is about 10%, about 20%, about 30%, about 40%, about 50%,about 60%, about 70%, about 80% or about 90% reduced when compared tothe AUC of a therapeutically effective amount of Compound 100A. In someembodiments, the AUC of a therapeutically effective amount of a compoundof Formula II that is not systemically absorbed (e.g., a compound ofFormula II that comprises a group L-K) is about 50% reduced whencompared to the AUC of a therapeutically effective amount of Compound100A. In other embodiments, the AUC of a therapeutically effectiveamount of a compound of Formula II that is not systemically absorbed(e.g., a compound of Formula II that comprises a group L-K) is about 75%reduced when compared to the AUC of a therapeutically effective amountof Compound 100A.

In some embodiments, the systemic exposure of a therapeuticallyeffective amount of a compound of Formula III, IIIA, IIIB or IIIC isreduced when compared to the systemic exposure of a therapeuticallyeffective amount of Compound 100C. In some embodiments, the AUC of atherapeutically effective amount of a compound of Formula III, IIIA,IIIB or IIIC is about 10%, about 20%, about 30%, about 40%, about 50%,about 60%, about 70%, about 80% or about 90% reduced when compared tothe AUC of a therapeutically effective amount of Compound 100C. In someembodiments, the AUC of a therapeutically effective amount of a compoundof Formula III, IIIA, IIIB or IIIC is about 50% reduced when compared tothe AUC of a therapeutically effective amount of Compound 100C. In otherembodiments, the AUC of a therapeutically effective amount of a compoundof Formula III, IIIA, IIIB or IIIC is about 75% reduced when compared tothe AUC of a therapeutically effective amount of Compound 100C.

In some embodiments, the systemic exposure of a therapeuticallyeffective amount of a compound of Formula IV that is not systemicallyabsorbed (e.g., a compound of Formula IV that comprises a group L-K) isreduced when compared to the systemic exposure of a therapeuticallyeffective amount of Compound 100A. In some embodiments, the AUC of atherapeutically effective amount of a compound of Formula IV that is notsystemically absorbed (e.g., a compound of Formula I that comprises agroup L-K) is about 10%, about 20%, about 30%, about 40%, about 50%,about 60%, about 70%, about 80% or about 90% reduced when compared tothe AUC of a therapeutically effective amount of Compound 100A. In someembodiments, the AUC of a therapeutically effective amount of a compoundof Formula IV that is not systemically absorbed (e.g., a compound ofFormula IV that comprises a group L-K) is about 50% reduced whencompared to the AUC of a therapeutically effective amount of Compound100A. In other embodiments, the AUC of a therapeutically effectiveamount of a compound of Formula IV that is not systemically absorbed(e.g., a compound of Formula IV that comprises a group L-K) is about 75%reduced when compared to the AUC of a therapeutically effective amountof Compound 100A.

In some embodiments, the systemic exposure of a therapeuticallyeffective amount of a compound of Formula V that is not systemicallyabsorbed (e.g., a compound of Formula V that comprises a group L-K) isreduced when compared to the systemic exposure of a therapeuticallyeffective amount of Compound 100A. In some embodiments, the AUC of atherapeutically effective amount of a compound of Formula V that is notsystemically absorbed (e.g., a compound of Formula V that comprises agroup L-K) is about 10%, about 20%, about 30%, about 40%, about 50%,about 60%, about 70%, about 80% or about 90% reduced when compared tothe AUC of a therapeutically effective amount of Compound 100A. In someembodiments, the AUC of a therapeutically effective amount of a compoundof Formula I that is not systemically absorbed (e.g., a compound ofFormula V that comprises a group L-K) is about 50% reduced when comparedto the AUC of a therapeutically effective amount of Compound 100A. Inother embodiments, the AUC of a therapeutically effective amount of acompound of Formula I that is not systemically absorbed (e.g., acompound of Formula V that comprises a group L-K) is about 75% reducedwhen compared to the AUC of a therapeutically effective amount ofCompound 100A.

In some embodiments, the systemic exposure of a therapeuticallyeffective amount of a compound of Formula VI or VID that is notsystemically absorbed (e.g., a compound of Formula VI or VID thatcomprises a group L-K) is reduced when compared to the systemic exposureof a therapeutically effective amount of Compound 100A. In someembodiments, the AUC of a therapeutically effective amount of a compoundof Formula VI or VID that is not systemically absorbed (e.g., a compoundof Formula VI or VID that comprises a group L-K) is about 10%, about20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80% orabout 90% reduced when compared to the AUC of a therapeuticallyeffective amount of Compound 100A. In some embodiments, the AUC of atherapeutically effective amount of a compound of Formula VI or VID thatis not systemically absorbed (e.g., a compound of Formula VI or VID thatcomprises a group L-K) is about 50% reduced when compared to the AUC ofa therapeutically effective amount of Compound 100A. In otherembodiments, the AUC of a therapeutically effective amount of a compoundof Formula I that is not systemically absorbed (e.g., a compound ofFormula VI or VID that comprises a group L-K) is about 75% reduced whencompared to the AUC of a therapeutically effective amount of Compound100A.

In certain embodiments, the Cmax of a therapeutically effective amountof any non-systemic ASBTI described herein (e.g., an ASBTI thatcomprises a group L-K) is at least 10%, at least 20%, at least 30%, atleast 40%, at least 50%, at least 60%, at least 70%, at least 80% or atleast 90% reduced when compared to the Cmax of any systemically absorbedASBTI (e.g. Compound 100A).

By way of example, the Cmax of a therapeutically effective amount of acompound of Formula III, IIIA, IIIB or IIIC is about 10%, about 20%,about 30%, about 40%, about 50%, about 60%, about 70%, about 80% orabout 90% reduced when compared to the Cmax of a therapeuticallyeffective amount of Compound 100C. In some embodiments, the Cmax of atherapeutically effective amount of a compound of Formula III, IIIA,IIIB or IIIC is about 25% reduced when compared to the Cmax of atherapeutically effective amount of Compound 100C. In certainembodiments, the Cmax of a therapeutically effective amount of acompound of III, IIIA or IIIB is about 50% reduced when compared to theCmax of a therapeutically effective amount of Compound 100C. In otherembodiments, the Cmax of a therapeutically effective amount of acompound of Formula III, IIIA, IIIB or IIIC is about 75% reduced whencompared to the Cmax of a therapeutically effective amount of Compound100C.

In certain embodiments, provided herein is a method of treating ametabolic disease or a condition associated with a metabolic diseasewith an ASBTI and/or enteroendocrine peptide secretion enhancing agentand an optional carrier. In some embodiments, provided herein aremethods of treating a metabolic disease comprising administering anycomposition as described herein. In some embodiments, provided herein isthe use of any of the pharmaceutical compositions described herein inpreparing a medicament for treating a metabolic disease. In specificembodiments, the metabolic disease is selected from, by way ofnon-limiting example, obesity, diabetes, necrotizing enterocolitis,gastritis, ulcerative colitis, inflammatory bowel disease, irritablebowel syndrome, gastroenteritis, radiation induced enteritis,chemotherapy induced enteritis, gastro-esophageal reflux disease (GERD),peptic ulcer, non-ulcer dyspepsia (NUD), celiac disease, intestinalceliac disease, gastrointestinal complications following bariatricsurgery, gastric carcinogenesis, gastric carcinogenesis followinggastric or bowel resection, and a combination thereof. In someembodiments, provided herein is a method of treating a conditionassociated with a metabolic disease. In certain embodiments, providedherein is the use of any of the pharmaceutical compositions describedherein in preparing a medicament for treating a condition associatedwith a metabolic disease. In some embodiments, conditions associatedwith a metabolic disease are selected from, by way of non-limitingexample, necrosis or damage of ileal architecture or ileal cells, weightgain, food intake, appetite, impaired glucose tolerance, a glucosemetabolic disorder, and insulin resistance. In certain embodiments,conditions associated with a metabolic disease are selected from, by wayof non-limiting example, acute coronary syndrome, hibernatingmyocardium, ventricular dysfunction, cardiac risk, post myocardialinfarction mortality, post-surgical catabolism, sepsis-relatedcatabolism, critical illness-related catabolism, post-surgicalmortality, sepsis-related mortality, critical illness-related mortality,critical illness-polyneuropathy, congestive heart failure, toxichypervolemia, renal failure, ischemia-reperfusion injury, mortality andmorbidity from stroke, mortality and morbidity from neurodegenerativedisease, neuropathy, inflammatory bowel disease, bowel mucosal injury,impaired bowel integrity, irritable bowel disease, osteopenia, and abone fracture or a bone disorder. It is to be understood that the term,“treating” includes controlling, suppressing, inhibiting, reducing thesymptoms of and/or preventing.

Furthermore, in certain embodiments, provided herein is a method ofdecreasing intraenterocyte bile acids, decreasing necrosis and/or damageto ileal architecture, decreasing appetite, decreasing food intake,and/or decreasing appetite by administering a therapeutically effectiveamount of any enteroendocrine peptide secretion enhancing agent orcomposition described herein. In some embodiments, provided herein is anenteroendocrine peptide secretion enhancing agent or a composition usedfor preparing a medicament for decreasing appetite, decreasing foodintake, and/or decreasing appetite.

In some embodiments, provided herein is a method of treating obesity,diabetes, or an inflammatory intestinal condition in an individualcomprising delivering to ileal, colon, and/or rectal L-cells of theindividual a therapeutically effective amount of any ASBTI and/orenteroendocrine peptide secretion enhancing agent described herein. Incertain embodiments, the therapeutically effective amount ofenteroendocrine peptide secretion enhancing agent stimulates oractivates the L-cells to which the enteroendocrine peptide secretionenhancing agent is administered.

FIG. 11 illustrates the reduction of food intake in response toadministration of anenteroendocrine peptide secretion enhancing agent.FIG. 11 illustrates that with increased amounts of enteroendocrinepeptide secretion enhancing agent administered to the distalgastrointestinal tract, decreased amounts of calories were consumed byhuman subjects.

In some embodiments, provided herein is a method of treating diabetes inan individual comprising delivering to ileal, colon, and/or rectalL-cells of an individual in need thereof a therapeutically effectiveamount of any enteroendocrine peptide secretion enhancing agentdescribed herein. In certain embodiments, provided herein is a method oftreating diabetes in an individual comprising delivering to ileum,colon, and/or rectum of an individual in need thereof a therapeuticallyeffective amount of any enteroendocrine peptide secretion enhancingagent described herein. In some embodiments, provided herein is a methodof elevating insulin levels in an individual comprising delivering toileum, colon, and/or rectum of an individual in need thereof (e.g., adiabetic individual) a therapeutically effective amount of anyenteroendocrine peptide secretion enhancing agent described herein. Incertain embodiments, provided herein is a method of reducing glucoselevels in an individual comprising delivering to ileum, colon, and/orrectum of an individual in need thereof (e.g., a diabetic individual) atherapeutically effective amount of any enteroendocrine peptidesecretion enhancing agent described herein. In some embodiments, thetherapeutically effective amount of enteroendocrine peptide secretionenhancing agent stimulates or activates the L-cells of the ileum, colon,and/or rectum to which the enteroendocrine peptide secretion enhancingagent is administered.

FIG. 12 illustrates the insulin response to administration of anenteroendocrine peptide secretion enhancing agent. FIG. 12 illustratesthat with increased amounts of enteroendocrine peptide secretionenhancing agent administered to the distal gastrointestinal tract, humandiabetics demonstrated increased levels of insulin. FIG. 13 illustratesthe incremental integrated response of insulin levels to administrationof an enteroendocrine peptide secretion enhancing agent.

FIG. 14 illustrates the glucose response to administration ofanenteroendocrine peptide secretion enhancing agent. FIG. 14 illustratesthat with increased amounts of enteroendocrine peptide secretionenhancing agent administered to the distal gastrointestinal tract, humandiabetics demonstrated decreased levels of glucose. FIG. 15 illustratesthe incremental integrated response of glucose levels to administrationof an enteroendocrine peptide secretion enhancing agent.

In some embodiments, provided herein is a method of elevating GLP-1,PYY, oxyntomodulin, insulin, or a combination thereof levels in anindividual comprising delivering to ileum, colon, and/or rectum of anindividual in need thereof (e.g., a diabetic individual) atherapeutically effective amount of any enteroendocrine peptidesecretion enhancing agent described herein. In some embodiments,provided herein is a method of elevating GLP-1, PYY, oxyntomodulin, andinsulin levels in an individual comprising delivering to ileum, colon,and/or rectum of an individual in need thereof (e.g., a diabeticindividual) a therapeutically effective amount of any enteroendocrinepeptide secretion enhancing agent described herein.

In some embodiments, provided herein is a method of preventing ortreating pancreatic or other cancers in an individual comprisingdelivering to ileal, colon, and/or rectal L-cells of an individual inneed thereof a therapeutically effective amount of any enteroendocrinepeptide secretion enhancing agent described herein. In certainembodiments, provided herein is a method of preventing or treatingpancreatic or other cancers in an individual comprising delivering toileum, colon, and/or rectum of an individual in need thereof atherapeutically effective amount of any enteroendocrine peptidesecretion enhancing agent described herein.

In some embodiments, the therapeutically effective amount ofenteroendocrine peptide secretion enhancing agent stimulates oractivates the L-cells of the ileum, colon, and/or rectum to which theenteroendocrine peptide secretion enhancing agent is administered. Othercancers include but are not limited to breast, rectal, colon and lungcancers.

In certain embodiments, the pharmaceutical composition administeredincludes a therapeutically effective amount of an enteroendocrinepeptide secretion enhancing agent, an absorption inhibitor and a carrier(e.g., an orally suitable carrier or a rectally suitable carrier,depending on the mode of intended administration). In certainembodiments, the pharmaceutical composition used or administeredcomprises an enteroendocrine peptide secretion enhancing agent, anabsorption inhibitor, a carrier, and one or more of a cholesterolabsorption inhibitor, an enteroendocrine peptide, a peptidase inhibitor,a spreading agent, and a wetting agent.

In a specific embodiment, the pharmaceutical composition used to preparea rectal dosage form or administered rectally comprises anenteroendocrine peptide secretion enhancing agent, an absorptioninhibitor, a rectally suitable carrier, an optional cholesterolabsorption inhibitor, an optional enteroendocrine peptide, an optionalpeptidase inhibitor, an optional spreading agent, and an optionalwetting agent. In certain embodiments, rectally administeredcompositions evokes an anorectal response. In specific embodiments, theanorectal response is an increase in secretion of one or moreenteroendocrine by cells (e.g., L-cells) in the colon and/or rectum(e.g., in the epithelial layer of the colon and/or rectum). In someembodiments, the anorectal response persists for at least 1, 2, 3, 4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24hours. In other embodiments the anorectal response persists for a periodbetween 24 hours and 48 hours, while in other embodiments the anorectalresponse persists for persists for a period greater than 48 hours.

In another specific embodiment, the pharmaceutical composition used toprepare an oral dosage form or administered orally comprises anenteroendocrine peptide secretion enhancing agent, an absorptioninhibitor, an orally suitable carrier, an optional cholesterolabsorption inhibitor, an optional enteroendocrine peptide, an optionalpeptidase inhibitor, an optional spreading agent, and an optionalwetting agent. In certain embodiments, the orally administeredcompositions evokes an anorectal response. In specific embodiments, theanorectal response is an increase in secretion of one or moreenteroendocrine by cells in the colon and/or rectum (e.g., in L-cellsthe epithelial layer of the colon and/or rectum). In some embodiments,the anorectal response persists for at least 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 hours. Inother embodiments the anorectal response persists for a period between24 hours and 48 hours, while in other embodiments the anorectal responsepersists for persists for a period greater than 48 hours.

Provided herein are methods for prevention and/or treatment ofcongestive heart failure, ventricular dysfunction, toxic hypervolemia,polycystic ovary syndrome, inflammatory bowel disease, impaired bowelintegrity, short bowel syndrome, gastritis, peptic ulcer, or irritablebowel disease comprising contacting the distal gastrointestinal tract(e.g., colon and/or rectum) of an individual in need thereof with anenteroendocrine peptide secretion enhancing agent and an optionalabsorption inhibitor. In some embodiments, the methods further compriseadministration of a DPP-IV inhibitor, an enteroendocrine peptideenhancing agent, a biguanide, an incretin mimetic, or GLP-1 or an analogthereof. Provided herein are methods for prevention and/or treatment ofradiation enteritis comprising contacting the distal gastrointestinaltract (e.g., colon and/or rectum) of an individual in need thereof withan enteroendocrine peptide secretion enhancing agent and an optionalabsorption inhibitor. In some embodiments, the methods further compriseadministration of a DPP-IV inhibitor, an enteroendocrine peptideenhancing agent, a biguanide, an incretin mimetic, or GLP-1 or an analogthereof. Provided in certain embodiments herein is a method of promotingregeneration of the gastrointestinal tract by administering to thedistal gastrointestinal tract (e.g., colon and/or rectum) of theindividual, a therapeutically effective amount of an enteroendocrinepeptide secretion enhancing agent and an optional absorption inhibitor.In some embodiments, the methods further comprise administration of aDPP-IV inhibitor, a TGR5 agonist, a biguanide, an incretin mimetic, orGLP-2 or an analog thereof. In some instances, radiation enteritis, oran inflammation of the intestine, is not only major complication ofcancer radiation therapy, but occur after any accidental and intentionalradiation exposure. In certain instances, GLP-2 secreted from L-cells(e.g., by administration thereto an enteroendocrine peptide secretionenhancing agent described herein) plays important role in regenerationof GI tract injuries.

Routes of Administration and Dosage

In some embodiments, the compositions described herein and thecompositions administered in the methods described herein are formulatedto enhance enteroendocrine peptide secretion and to evoke an anorectalresponse. In certain embodiments, the compositions described herein areformulated for rectal or oral administration. In some embodiments, suchformulations are administered rectally or orally, respectively. In someembodiments, the compositions described herein are combined with adevice for local delivery of the compositions to the rectum and/or colon(sigmoid colon, transverse colon, or ascending colon). In certainembodiments, for rectal administration the composition described hereinare formulated as enemas, rectal gels, rectal foams, rectal aerosols,suppositories, jelly suppositories, or retention enemas. In someembodiments, for oral administration the compositions described hereinare formulated for oral administration and enteric delivery to thecolon.

In certain embodiments, the compositions or methods described herein arenon-systemic. In some embodiments, compositions described herein deliverthe enteroendocrine peptide secretion enhancing agent to the distalileum, colon, and/or rectum and not systemically (e.g., a substantialportion of the enteroendocrine peptide secretion enhancing agent is notsystemically absorbed). In some embodiments, oral compositions describedherein deliver the enteroendocrine peptide secretion enhancing agent tothe distal ileum, colon, and/or rectum and not systemically (e.g., asubstantial portion of the enteroendocrine peptide secretion enhancingagent is not systemically absorbed). In some embodiments, rectalcompositions described herein deliver the enteroendocrine peptidesecretion enhancing agent to the distal ileum, colon, and/or rectum andnot systemically (e.g., a substantial portion of the enteroendocrinepeptide secretion enhancing agent is not systemically absorbed). Incertain embodiments, non-systemic compositions described herein deliverless than 90% w/w of the enteroendocrine peptide secretion enhancingagent systemically. In certain embodiments, non-systemic compositionsdescribed herein deliver less than 80% w/w of the enteroendocrinepeptide secretion enhancing agent systemically. In certain embodiments,non-systemic compositions described herein deliver less than 70% w/w ofthe enteroendocrine peptide secretion enhancing agent systemically. Incertain embodiments, non-systemic compositions described herein deliverless than 60% w/w of the enteroendocrine peptide secretion enhancingagent systemically. In certain embodiments, non-systemic compositionsdescribed herein deliver less than 50% w/w of the enteroendocrinepeptide secretion enhancing agent systemically. In certain embodiments,non-systemic compositions described herein deliver less than 40% w/w ofthe enteroendocrine peptide secretion enhancing agent systemically. Incertain embodiments, non-systemic compositions described herein deliverless than 30% w/w of the enteroendocrine peptide secretion enhancingagent systemically. In certain embodiments, non-systemic compositionsdescribed herein deliver less than 25% w/w of the enteroendocrinepeptide secretion enhancing agent systemically. In certain embodiments,non-systemic compositions described herein deliver less than 20% w/w ofthe enteroendocrine peptide secretion enhancing agent systemically. Incertain embodiments, non-systemic compositions described herein deliverless than 15% w/w of the enteroendocrine peptide secretion enhancingagent systemically. In certain embodiments, non-systemic compositionsdescribed herein deliver less than 10% w/w of the enteroendocrinepeptide secretion enhancing agent systemically. In certain embodiments,non-systemic compositions described herein deliver less than 5% w/w ofthe enteroendocrine peptide secretion enhancing agent systemically. Insome embodiments, systemic absorption is determined in any suitablemanner, including the total circulating amount, the amount cleared afteradministration, or the like.

In certain embodiments, the compositions and/or formulations describedherein are administered at least once a day. In certain embodiments, theformulations containing the enteroendocrine peptide secretion enhancingagents are administered at least twice a day, while in other embodimentsthe formulations containing the enteroendocrine peptide secretionenhancing agents are administered at least three times a day. In certainembodiments, the formulations containing the enteroendocrine peptidesecretion enhancing agents are administered up to five times a day. Itis to be understood that in certain embodiments, the dosage regimen ofcomposition containing the enteroendocrine peptide secretion enhancingagents described herein to is determined by considering various factorssuch as the patient's age, sex, and diet.

The concentration of the enteroendocrine peptide secretion enhancingagents administered in the formulations described herein ranges fromabout 1 mM to about 1 M. In certain embodiments the concentration of theenteroendocrine peptide secretion enhancing agents administered in theformulations described herein ranges from about 1 mM to about 750 mM. Incertain embodiments the concentration of the enteroendocrine peptidesecretion enhancing agents administered in the formulations describedherein ranges from about 1 mM to about 500 mM. In certain embodimentsthe concentration of the enteroendocrine peptide secretion enhancingagents administered in the formulations described herein ranges fromabout 5 mM to about 500 mM. In certain embodiments the concentration ofthe enteroendocrine peptide secretion enhancing agents administered inthe formulations described herein ranges from about 10 mM to about 500mM. In certain embodiments the concentration of the enteroendocrinepeptide secretion enhancing agents administered in the formulationsdescribed herein ranges from about 25 mM to about 500 mM. In certainembodiments the concentration of the enteroendocrine peptide secretionenhancing agents administered in the formulations described hereinranges from about 50 mM to about 500 mM. In certain embodiments theconcentration of the enteroendocrine peptide secretion enhancing agentsadministered in the formulations described herein ranges from about 100mM to about 500 mM. In certain embodiments the concentration of theenteroendocrine peptide secretion enhancing agents administered in theformulations described herein ranges from about 200 mM to about 500 mM.

In certain embodiments, any composition described herein comprises atherapeutically effective amount (e.g., to treat obesity and/ordiabetes) of an enteroendocrine peptide secretion enhancing agent (e.g.,bile acid). In some embodiments, compositions described herein compriseor methods described herein comprise administering about 0.01 mg toabout 10 g of an enteroendocrine peptide secretion enhancing agent(e.g., bile acid). In certain embodiments, a composition describedherein comprises or a method described herein comprises administeringabout 0.1 mg to about 500 mg of an enteroendocrine peptide secretionenhancing agent (e.g., bile acid). In certain embodiments, a compositiondescribed herein comprises or a method described herein comprisesadministering about 0.1 mg to about 100 mg of an enteroendocrine peptidesecretion enhancing agent (e.g., bile acid). In certain embodiments, acomposition described herein comprises or a method described hereincomprises administering about 0.1 mg to about 50 mg of anenteroendocrine peptide secretion enhancing agent (e.g., bile acid). Incertain embodiments, a composition described herein comprises or amethod described herein comprises administering about 0.1 mg to about 10mg of an enteroendocrine peptide secretion enhancing agent (e.g., bileacid). In certain embodiments, a composition described herein comprisesor a method described herein comprises administering about 0.5 mg toabout 10 mg of an enteroendocrine peptide secretion enhancing agent(e.g., bile acid). In some embodiments, compositions described hereincomprise or methods described herein comprise administering about 0.1mmol to about 1 mol of an enteroendocrine peptide secretion enhancingagent (e.g., bile acid). In certain embodiments, a composition describedherein comprises or a method described herein comprises administeringabout 0.01 mmol to about 500 mmol of an enteroendocrine peptidesecretion enhancing agent (e.g., bile acid). In certain embodiments, acomposition described herein comprises or a method described hereincomprises administering about 0.1 mmol to about 100 mmol of anenteroendocrine peptide secretion enhancing agent (e.g., bile acid). Incertain embodiments, a composition described herein comprises or amethod described herein comprises administering about 0.5 mmol to about30 mmol of an enteroendocrine peptide secretion enhancing agent (e.g.,bile acid). In certain embodiments, a composition described hereincomprises or a method described herein comprises administering about 0.5mmol to about 20 mmol of an enteroendocrine peptide secretion enhancingagent (e.g., bile acid). In certain embodiments, a composition describedherein comprises or a method described herein comprises administeringabout 1 mmol to about 10 mmol of an enteroendocrine peptide secretionenhancing agent (e.g., bile acid). In certain embodiments, a compositiondescribed herein comprises or a method described herein comprisesadministering about 0.01 mmol to about 5 mmol of an enteroendocrinepeptide secretion enhancing agent (e.g., bile acid). In certainembodiments, a composition described herein comprises or a methoddescribed herein comprises administering about 0.1 mmol to about 1 mmolof an enteroendocrine peptide secretion enhancing agent (e.g., bileacid). In various embodiments, certain enteroendocrine peptide secretionenhancing agents (e.g., bile acids) have different potencies and dosingis optionally adjusted accordingly. For example, the investigation inTGR5-transfected CHO cells of TGR5 agonist potency of natural bile acidsindicates the following rank of potency: Lithocholic acid(LCA)>deoxycholic acid (DCA)>murocholic acid (Muro-CA)>lagodeoxycholicacid (lago-DCA)>chenodeoxycholic (CDCA)>cholic acid (CA)>hyodeoxycholicacid (HDCA>ursodeoxycholic acid (UDCA); and assays on TGR5-transfectedCHO cells demonstrate that EC₅₀ (in μM) for UDCA was 36.4, TauroCA (TCA)4.95 and LCA 0.58.

In certain embodiments, by targeting the distal gastrointestinal tract(e.g., distal ileum, colon, and/or rectum), compositions and methodsdescribed herein provide efficacy (e.g., in reducing food intake,treating obesity, treating diabetes) with a reduced dose ofenteroendocrine peptide secretion enhancing agent (e.g., as compared toan oral dose that does not target the distal gastrointestinal tract).

Rectal Administration Formulations

The pharmaceutical compositions described herein for the non-systemicdelivery of enteroendocrine peptide secretion enhancing agents to therectum and/or colon are formulated for rectal administration as rectalenemas, rectal foams, rectal gels, and rectal suppositories. Thecomponents of such formulations are described herein. It is to beunderstood that as used herein, pharmaceutical compositions andcompositions are or comprise the formulations as described herein.

Rectal Enemas

In certain embodiments, the compositions described herein are formulatedas rectal enema formulations for non-systemic delivery ofenteroendocrine peptide secretion enhancing agents. In certainembodiments, such rectal enemas are formulated as a solution, aqueoussuspension or emulsion. In some embodiments, solution enemas contain acarrier vehicle, an enteroendocrine peptide secretion enhancing agent,an absorption inhibitor (e.g., of the enteroendocrine peptide secretionenhancing agent across the rectal or colonic mucosa), and one or more ofthe following: a solubilizer, a preservative, a chelating agent, abuffer for pH regulation, and a thickener. In certain embodiments,rectal enemas are formulated as an emulsion or aqueous suspensioncontaining a carrier vehicle, at least one enteroendocrine peptidesecretion enhancing agent, at least one agent for inhibiting absorptionof the enteroendocrine peptide secretion enhancing agent across therectal or colonic mucosa, and one or more of the following: apreservative, a chelating agent, a buffer for pH regulation, asolubilizer, a thickener, and an emulsifier/surfactant.

In certain embodiments, rectal enemas are formulated such that aenteroendocrine peptide secretion enhancing agent is dissolved ordispersed in a suitable flowable carrier vehicle, including but notlimited to water, alcohol or an aqueous-alcoholic mixture. In certainembodiments, the carrier vehicle is thickened with natural or syntheticthickeners. In further embodiments the rectal enema formulations alsocontain a lubricant.

In some embodiments, unit dosages of such enema formulations areadministered from prefilled bags or syringes.

In certain embodiments, the volume of enema administered using suchrectal enema formulations is a volume suitable for achieving a desiredresult, e.g., from about 10 mL to about 1000 mL. In certain embodiments,the volume of enema administered using such rectal enema formulations isfrom about 10 mL to about 900 mL. In certain embodiments, the volume ofenema administered using such rectal enema formulations is from about 10mL to about 800 mL. In certain embodiments, the volume of enemaadministered using such rectal enema formulations is from about 10 mL toabout 700 mL. In certain embodiments, the volume of enema administeredusing such rectal enema formulations is from about 10 mL to about 600mL. In certain embodiments, the volume of enema administered using suchrectal enema formulations is from about 10 mL to about 500 mL. Incertain embodiments, the volume of enema administered using such rectalenema formulations is from about 10 mL to about 400 mL. In certainembodiments, the volume of enema administered using such rectal enemaformulations is from about 10 mL to about 300 mL. In certainembodiments, the volume of enema administered using such rectal enemaformulations is from about 10 mL to about 200 mL. In certainembodiments, the volume of enema administered using such rectal enemaformulations is from about 10 mL to about 100 mL. In some embodiments,such enemas may have a volume of less than 1 L, less than 900 mL, lessthan 700 mL, less than 600 mL, less than 500 mL, less than 250 mL, lessthan 100 mL, less than 30 mL, less than 10 mL, less than 3 mL, or thelike.

Rectal Foams

In certain instances, leakage is a problem associated with enemas. Assuch, it is often desirable or necessary for patients to lie down duringadministration of enemas. In some embodiments, rectal administrationusing foams overcomes the problem of leakage from the rectum followingadministration.

In certain embodiments, the pharmaceutical compositions are formulatedas rectal foams. In some embodiments, rectal foams are used for therectal administration and for local or non-systemic delivery ofenteroendocrine peptide secretion enhancing agents to the rectum and/orcolon. Such rectal foams formulations contain an enteroendocrine peptidesecretion enhancing agent dissolved or suspended in a liquid carriervehicle, an absorption inhibitor (e.g., of the enteroendocrine peptidesecretion enhancing agent across the rectal or colonic mucosa), asurfactant/emulsifier with foaming properties and a propellant (e.g., apropellant gas). In certain embodiments, rectal foam formulations alsocontain one or more of the following: a suspending/solubilizing agent, athickener, a preservative, a chelating agent, a buffer, an antioxidant,a tonicity modifiers, and a spreading agent. In certain embodiments,surfactants/emulsifiers include, by way of non-limiting example,non-ionic surfactants, anionic surfactants, cationic surfactants, andcombinations thereof.

In certain embodiments, rectal foam formulations are filled inpressurized containers prior to rectal administration. In certainembodiments the pressurized container is a can. In certain embodiments,propellants used herein include, by way of non-limiting example,hydrocarbons (such as isobutane, N-butane or propane), fluorocarbons(e.g. dichlorodifluoromethane and dichlorotetrafluoroethane),chlorofluorocarbons, dimethyl ether, hydrofluorocarbons, compressedgases, freon (such as freon 12, freon 114), hydrochlorofluorocarbons,hydrofluorocarbons or mixtures thereof.

In some embodiments, the maximum amount of propellant used is determinedby its miscibility with other components in the composition to form amixture, such as a homogeneous mixture. In certain embodiments, theminimal level of propellant used in the composition is determined by thedesired foam characteristics, and its ability to substantially orcompletely evacuate the container.

In some embodiments, the propellant concentration used in such rectalfoam formulations is about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%,12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 50%,55% to about 60% (w/w).

In certain embodiments, rectal foams are formed upon rectaladministration, wherein the dispensing valve of the can allows rapidexpansion of the propellant, triggering the foaming action of thesurfactant and resulting foam forms within the rectum and colon. Inother embodiments, the rectal foams used for rectal administration ofthe compositions described herein are formed within the dispensingcontainer prior to rectal administration.

The distance the foam can reach within the colon and rectum iscontrolled by controlling the foam propelling properties by varying thetype and quantity of propellant used. The volume of foam administeredusing such rectal foam formulations is from about 10 mL to about 1000mL. In certain embodiments, the volume of a composition described herein(e.g., a foam) described herein or used in a method described herein(e.g., a foam, enema, or gel) is from about 10 mL to about 900 mL. Incertain embodiments, the volume of a composition described herein (e.g.,a foam) described herein or used in a method described herein (e.g., afoam, enema, or gel) is from about 10 mL to about 800 mL. In certainembodiments, the volume of a composition described herein (e.g., a foam)described herein or used in a method described herein (e.g., a foam,enema, or gel) is from about 10 mL to about 700 mL. In certainembodiments, the volume of a composition described herein (e.g., a foam)described herein or used in a method described herein (e.g., a foam,enema, or gel) is from about 10 mL to about 600 mL. In certainembodiments, the volume of a composition described herein (e.g., a foam)described herein or used in a method described herein (e.g., a foam,enema, or gel) is from about 10 mL to about 500 mL. In certainembodiments, the volume of a composition described herein (e.g., a foam)described herein or used in a method described herein (e.g., a foam,enema, or gel) is from about 10 mL to about 400 mL. In certainembodiments, the volume of a composition described herein (e.g., a foam)described herein or used in a method described herein (e.g., a foam,enema, or gel) is from about 10 mL to about 300 mL. In certainembodiments, the volume of a composition described herein (e.g., a foam)described herein or used in a method described herein (e.g., a foam,enema, or gel) is from about 10 mL to about 200 mL. In certainembodiments, the volume of a composition described herein (e.g., a foam)described herein or used in a method described herein (e.g., a foam,enema, or gel) is from about 10 mL to about 100 mL. In specificembodiments, the volume of a composition described herein (e.g., a foam)described herein or used in a method described herein (e.g., a foam,enema, or gel) is about 20 mL to about 60 mL, about 20 mL, about 40 mL,or about 60 mL.

Rectal Gels

In some embodiments, the pharmaceutical compositions described hereinare formulated as rectal gels. In certain embodiments, the rectal gelsare suitable for the regional or local non-systemic administration ofone or more enteroendocrine peptide secretion enhancing agents to therectum and/or colon. In some embodiments, rectal gel formulationscontain at least one enteroendocrine peptide secretion enhancing agentdissolved or suspended in a solvent/liquid carrier vehicle, anabsorption inhibitor (e.g., of the enteroendocrine peptide secretionenhancing agent across the rectal or colonic mucosa) and at least onethickening agents. In certain embodiments such rectal gel formulationsalso contain one or more of the following: a buffering agent(s), apreservative(s), and an antioxidant(s).

In certain embodiments, rectal gels have gel-like consistencies but aresufficiently flowable so as to be capable of local or regionaladministration through a catheter, needle, syringe, or other comparablemeans of local or regional administration.

In some embodiments, the concentration of a thickener used in a rectalgel formulation is in an amount or concentration suitable to achieve adesired thickness or viscosity, e.g., from about 0.05% to about 10% byweight. In certain embodiments, the concentration of the thickener usedin such rectal gel formulations ranges from about 0.05% to about 8% byweight. In certain embodiments, the concentration of the thickener usedin such rectal gel formulations ranges from about 0.05% to about 7% byweight. In certain embodiments, the concentration of the thickener usedin such rectal gel formulations ranges from about 0.05% to about 6% byweight. In certain embodiments, the concentration of the thickener usedin such rectal gel formulations ranges from about 0.05% to about 5% byweight. In certain embodiments, the concentration of the thickener usedin such rectal gel formulations ranges from about 0.05% to about 4% byweight. In certain embodiments, the concentration of the thickener usedin such rectal gel formulations ranges from about 0.05% to about 3% byweight. In certain embodiments, the concentration of the thickener usedin such rectal gel formulations ranges from about 0.05% to about 2% byweight. In certain embodiments, the concentration of the thickener usedin such rectal gel formulations ranges from about 0.05% to about 1% byweight. In certain embodiments the rectal gel formulation includesmethyl cellulose having a concentration from about 0.05% to about 2%,while in other embodiments the rectal gel formulation includes methylcellulose having a concentration of about 1%.

In some embodiments, the any formulation described herein (e.g., arectal gel formulation) has a viscosity ranging from about 500 to about50,000 centipoise (cP) at 25 C. In certain embodiments, the viscosity ofthe formulation described herein is from about 500 to about 40,000centipoise (cP) at 25 C. In certain embodiments, the viscosity of theformulation described herein is from about 500 to about 30,000centipoise (cP) at 25 C. In certain embodiments, the viscosity of theformulation described herein is from about 500 to about 20,000centipoise (cP) at 25 C. In certain embodiments, the viscosity of theformulation described herein is from about 500 to about 10,000centipoise (cP) at 25 C. In some embodiments, the formulation has afinal viscosity of less than about 40,000 centipoises (cP), 20,000 cP,15,000 cP, or 10,000 cP at 25 C. In some embodiments, the formulationhas a viscosity of about 5,000 cP, 6,000 cP, 7,000 cP, 8,000 cP, 9,000cP, 10,000 cP, 12,000 cP, 15,000 cP, 18,000 cP, 20,000 cP, 25,000 cP,30,000 cP, 35,000 cP, or 40,000 cP at 25 C. In some embodiments, theformulation has a viscosity of about 1,000-20,000 cP, 5,000-15,000 cP,6,000-12,000 cP, 7,000-10,000, 500-3500 cP, 500-300 cP, 1,000-2,000 cP,or about 1,500 cP at 25 C. In specific embodiments, the formulation hasa viscosity of 1,000 cP to about 2,500 cP, or about 1,500 cP at 25 C. Incertain embodiments, the amount of thickener used in a compositiondescribed herein is sufficient to achieve a viscosity as describedherein. FIG. 4 illustrates the affect of the viscosity of a formulationdescribed herein on the food intake of a subject.

In some embodiments, unit dosages of such rectal gel formulations areadministered from prefilled bags or syringes.

Rectal Suppositories

In some embodiments, the pharmaceutical compositions described hereinare also formulated as a suppository. In certain embodiments,suppositories are formulated for the regional or local non-systemicadministration of one or more enteroendocrine peptide secretionenhancing agents to the rectum and/or colon.

In some embodiments, rectal suppository formulations contain aenteroendocrine peptide secretion enhancing agent, an absorptioninhibitor (e.g., of the enteroendocrine peptide secretion enhancingagent across the rectal or colonic mucosa) and at least onepharmaceutically acceptable suppository base. In some embodiments,suppository formulation are prepared by combining an enteroendocrinepeptide secretion enhancing agent with a pharmaceutically acceptablesuppository base, melted, poured into a mould or moulds and cooled.

In certain embodiments, pharmaceutically acceptable suppository basesinclude, by way of non-limiting example, cocoa butter, beeswax,esterified fatty acids, glycerinated gelatin, semisynthetic glyceridesof vegetable saturated fatty acids, polyethylene glycols, Witepsol, andpolyoxyethylene sorbitan fatty acid esters.

In certain embodiments, the suppository formulations used to deliver oneor more enteroendocrine peptide secretion enhancing agents to the rectumand/or colon also contain one or more of the following: bufferingagents, preservatives, antioxidants, surfactants, and thickeners.

In some embodiments, suppositories contain from 0.5 to 10 mg of anenteroendocrine peptide secretion enhancing agent. In specificembodiments, suppositories contain from 1 to 5 mg of an enteroendocrinepeptide secretion enhancing agent.

Components Used in Rectal Delivery/Administration Formulations

In certain embodiments, liquid carrier vehicles in the compositionsand/or formulations described herein include, by way of non-limitingexample, purified water, propylene glycol, polyethyleneglycol, ethanol,1-propanol, 2-propanol, 1-propen-3-ol (allyl alcohol), propylene glycol,glycerol, 2-methyl-2-propanol, formamide, methyl formamide, dimethylformamide, ethyl formamide, diethyl formamide, acetamide, methylacetamide, dimethyl acetamide, ethyl acetamide, diethyl acetamide,2-pyrrolidone, N-methyl-2-pyrrolidone, N-ethyl-2-pyrrolidone,tetramethyl urea, 1,3-dimethyl-2-imidazolidinone, propylene carbonate,1,2-butylene carbonate, 2,3-butylene carbonate, dimethyl sulfoxide,diethyl sulfoxide, hexamethyl phosphoramide, pyruvic aldehydedimethylacetal, dimethylisosorbide and combinations thereof.

In some embodiments, stabilizers used in compositions and/orformulations described herein include, but are not limited to, partialglycerides of polyoxyethylenic saturated fatty acids.

In certain embodiments, surfactants/emulsifiers used in the compositionsand/or formulations described herein include, by way of non-limitingexample, mixtures of cetostearylic alcohol with sorbitan esterified withpolyoxyethylenic fatty acids, polyoxyethylene fatty ethers,polyoxyethylene fatty esters, fatty acids, sulfated fatty acids,phosphated fatty acids, sulfosuccinates, amphoteric surfactants,non-ionic poloxamers, non-ionic meroxapols, petroleum derivatives,aliphatic amines, polysiloxane derivatives, sorbitan fatty acid esters,laureth-4, PEG-2 dilaurate, stearic acid, sodium lauryl sulfate, dioctylsodium sulfosuccinate, cocoamphopropionate, poloxamer 188, meroxapol258, triethanolamine, dimethicone, polysorbate 60, sorbitanmonostearate, pharmaceutically acceptable salts thereof, andcombinations thereof.

In some embodiments, non-ionic surfactants used in compositions and/orformulations described herein include, by way of non-limiting example,phospholipids, alkyl poly(ethylene oxide), poloxamers, polysorbates,sodium dioctyl sulfosuccinate, Brij™-30 (Laureth-4), Brij™-58(Ceteth-20) and Brij™-78 (Steareth-20), Brij™-721 (Steareth-21),Crillet-1 (Polysorbate 20), Crillet-2 (Polysorbate 40), Crillet-3(Polysorbate 60), Crillet 45 (Polysorbate 80), Myrj-52 (PEG-40Stearate), Myrj-53 (PEG-50 Stearate), Pluronic™ F77 (Poloxamer 217),Pluronic™ F87 (Poloxamer 237), Pluronic™ F98 (Poloxamer 288), Pluronic™L62 (Poloxamer 182), Pluronic™ L64 (Poloxamer 184), Pluronic™ F68(Poloxamer 188), Pluronic™ L81 (Poloxamer 231), Pluronic™ L92 (Poloxamer282), Pluronic™ L101 (Poloxamer 331), Pluronic™ P103 (Poloxamer 333),Pluracare™ F 108 NF (Poloxamer 338), and Pluracare™ F 127 NF (Poloxamer407) and combinations thereof. Pluronic™ polymers are commerciallypurchasable from BASF, USA and Germany.

In certain embodiments, anionic surfactants used in compositions and/orformulations described herein include, by way of non-limiting example,sodium laurylsulphate, sodium dodecyl sulfate (SDS), ammonium laurylsulfate, alkyl sulfate salts, alkyl benzene sulfonate, and combinationsthereof.

In some embodiments, the cationic surfactants used in compositionsand/or formulations described herein include, by way of non-limitingexample, benzalkonium chloride, benzethonium chloride, cetyltrimethylammonium bromide, hexadecyl trimethyl ammonium bromide, otheralkyltrimethylammonium salts, cetylpyridinium chloride, polyethoxylatedtallow and combinations thereof.

In certain embodiments, the thickeners used i in compositions and/orformulations described herein include, by way of non-limiting example,natural polysaccharides, semi-synthetic polymers, synthetic polymers,and combinations thereof. Natural polysaccharides include, by way ofnon-limiting example, acacia, agar, alginates, carrageenan, guar,arabic, tragacanth gum, pectins, dextran, gellan and xanthan gums.

Semi-synthetic polymers include, by way of non-limiting example,cellulose esters, modified starches, modified celluloses,carboxymethylcellulose, methyl cellulose, ethyl cellulose, hydroxyethylcellulose, hydroxypropyl cellulose and hydroxypropyl methylcellulose.Synthetic polymers include, by way of non-limiting example,polyoxyalkylenes, polyvinyl alcohol, polyacrylamide, polyacrylates,carboxypolymethylene (carbomer), polyvinylpyrrolidone (povidones),polyvinylacetate, polyethylene glycols and poloxamer. Other thickenersinclude, by way of nonlimiting example, polyoxyethyleneglycolisostearate, cetyl alcohol, Polyglycol 300 isostearate, propyleneglycol,collagen, gelatin, and fatty acids (e.g., lauric acid, myristic acid,palmitic acid, stearic acid, palmitoleic acid, linoleic acid, linolenicacid, oleic acid and the like).

In some embodiments, chelating agents used in the compositions and/orformulations described herein include, by way of non-limiting example,ethylenediaminetetraacetic acid (EDTA) or salts thereof, phosphates andcombinations thereof.

In some embodiments, the concentration of the chelating agent or agentsused in the rectal formulations described herein is a suitableconcentration, e.g., about 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.4%, or 0.5%(w/v).

In some embodiments, preservatives used in compositions and/orformulations described herein include, by way of non-limiting example,parabens, ascorbyl palmitate, benzoic acid, butylated hydroxyanisole,butylated hydroxytoluene, chlorobutanol, ethylenediamine, ethylparaben,methylparaben, butyl paraben, propylparaben, monothioglycerol, phenol,phenylethyl alcohol, propylparaben, sodium benzoate, sodium propionate,sodium formaldehyde sulfoxylate, sodium metabisulfite, sorbic acid,sulfur dioxide, maleic acid, propyl gallate, benzalkonium chloride,benzethonium chloride, benzyl alcohol, chlorhexidine acetate,chlorhexidine gluconate, sorbic acid, potassium sorbitol, chlorbutanol,phenoxyethanol, cetylpyridinium chloride, phenylmercuric nitrate,thimerosol, and combinations thereof.

In certain embodiments, antioxidants used in compositions and/orformulations described herein include, by way of non-limiting example,ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylatedhydroxytoluene, hypophosphorous acid, monothioglycerol, propyl gallate,sodium ascorbate, sodium sulfite, sodium bisulfite, sodium formaldehydesulfoxylate, potassium metabisulphite, sodium metabisulfite, oxygen,quinones, t-butyl hydroquinone, erythorbic acid, olive (olea eurpaea)oil, pentasodium penetetate, pentetic acid, tocopheryl, tocopherylacetate and combinations thereof.

In some embodiments, concentration of the antioxidant or antioxidantsused in the rectal formulations described herein is sufficient toachieve a desired result, e.g., about 0.1%, 0.15%, 0.2%, 0.25%, 0.3%,0.4%, or 0.5% (w/v).

The lubricating agents used in compositions and/or formulationsdescribed herein include, by way of non-limiting example, natural orsynthetic fat or oil (e.g., a tris-fatty acid glycerate and the like).In some embodiments, lubricating agents include, by way of non-limitingexample, glycerin (also called glycerine, glycerol, 1,2,3-propanetriol,and trihydroxypropane), polyethylene glycols (PEGs), polypropyleneglycol, polyisobutene, polyethylene oxide, behenic acid, behenylalcohol, sorbitol, mannitol, lactose, polydimethylsiloxane andcombinations thereof.

In certain embodiments, mucoadhesive and/or bioadhesive polymers areused in the compositions and/or formulations described herein as agentsfor inhibiting absorption of the enteroendocrine peptide secretionenhancing agent across the rectal or colonic mucosa. Bioadhesive ormucoadhesive polymers include, by way of non-limiting example,hydroxypropyl cellulose, polyethylene oxide homopolymers, polyvinylether-maleic acid copolymers, methyl cellulose, ethyl cellulose, propylcellulose, hydroxyethyl cellulose, hydroxypropyl cellulose,hydroxypropylmethyl cellulose, carboxymethylcellulose, polycarbophil,polyvinylpyrrolidone, carbopol, polyurethanes, polyethyleneoxide-polypropyline oxide copolymers, sodium carboxymethyl cellulose,polyethylene, polypropylene, lectins, xanthan gum, alginates, sodiumalginate, polyacrylic acid, chitosan, hyaluronic acid and esterderivatives thereof, vinyl acetate homopolymer, calcium polycarbophil,gelatin, natural gums, karaya, tragacanth, algin, chitosan, starches,pectins, and combinations thereof.

In some embodiments, buffers/pH adjusting agents used in compositionsand/or formulations described herein include, by way of non-limitingexample, phosphoric acid, monobasic sodium or potassium phosphate,triethanolamine (TRIS), BICINE, HEPES, Trizma, glycine, histidine,arginine, lysine, asparagine, aspartic acid, glutamine, glutamic acid,carbonate, bicarbonate, potassium metaphosphate, potassium phosphate,monobasic sodium acetate, acetic acid, acetate, citric acid, sodiumcitrate anhydrous, sodium citrate dihydrate and combinations thereof. Incertain embodiments, an acid or a base is added to adjust the pH.Suitable acids or bases include, by way of non-limiting example, HCL,NaOH and KOH.

In certain embodiments, concentration of the buffering agent or agentsused in the rectal formulations described herein is sufficient toachieve or maintain a physiologically desirable pH, e.g., about 0.1%,0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.8%, 0.9%, or 1.0% (w/w).

The tonicity modifiers used in compositions and/or formulationsdescribed herein include, by way of non-limiting example o, sodiumchloride, potassium chloride, sodium phosphate, mannitol, sorbitol orglucose.

Devices

In certain aspects of the methods and pharmaceutical compositionsdescribed herein, a device is used for rectal administration of thecompositions and/or formulations described herein (e.g., the rectalgels, rectal foams, enemas and suppositories described herein). Incertain embodiments, rectal gels or rectal enemas are administered usinga bag or a syringe, while rectal foams are administered using apressurized container.

In certain embodiments, a perfusion system is used to rectallyadminister the pharmaceutical compositions and/or formulations describedherein. In some embodiments, the system comprises a tube surrounded by asemi-permeable membrane is rectally inserted and a solution containing acomposition described herein is pumped into the membrane. In certainembodiments, the membrane expands to contact the rectal and/or colonwalls, wherein the enterendocrine peptide secretion enhancing agentsperfuse from the inside of the membrane to the outside. In certainembodiments, the solution is re-circulated as a continuous perfusionsystem.

Oral Administration for Colonic Delivery

In certain aspects, the composition or formulation containing one ormore enteroendocrine peptide secretion enhancing agents is orallyadministered for local delivery of an ASBTI, a TGR5 agonist, or anenteroendocrine peptide secretion enhancing agent to the colon and/orrectum. Unit dosage forms of such compositions include a pill, tablet orcapsules formulated for enteric delivery to colon. In certainembodiments, such pills, tablets or capsule contain the compositionsdescribed herein entrapped or embedded in microspheres. In someembodiments, microspheres include, by way of non-limiting example,chitosan microcores HPMC capsules and cellulose acetate butyrate (CAB)microspheres. In certain embodiments, oral dosage forms are preparedusing conventional methods known to those in the field of pharmaceuticalformulation. For example, in certain embodiments, tablets aremanufactured using standard tablet processing procedures and equipment.An exemplary method for forming tablets is by direct compression of apowdered, crystalline or granular composition containing the activeagent(s), alone or in combination with one or more carriers, additives,or the like. In alternative embodiments, tablets are prepared usingwet-granulation or dry-granulation processes. In some embodiments,tablets are molded rather than compressed, starting with a moist orotherwise tractable material.

In certain embodiments, tablets prepared for oral administration containvarious excipients, including, by way of non-limiting example, binders,diluents, lubricants, disintegrants, fillers, stabilizers, surfactants,preservatives, coloring agents, flavoring agents and the like. In someembodiments, binders are used to impart cohesive qualities to a tablet,ensuring that the tablet remains intact after compression. Suitablebinder materials include, by way of non-limiting example, starch(including corn starch and pregelatinized starch), gelatin, sugars(including sucrose, glucose, dextrose and lactose), polyethylene glycol,propylene glycol, waxes, and natural and synthetic gums, e.g., acaciasodium alginate, polyvinylpyrrolidone, cellulosic polymers (includinghydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose, ethyl cellulose, hydroxyethyl cellulose, and the like),Veegum, and combinations thereof. In certain embodiments, diluents areutilized to increase the bulk of the tablet so that a practical sizetablet is provided. Suitable diluents include, by way of non-limitingexample, dicalcium phosphate, calcium sulfate, lactose, cellulose,kaolin, mannitol, sodium chloride, dry starch, powdered sugar andcombinations thereof. In certain embodiments, lubricants are used tofacilitate tablet manufacture; examples of suitable lubricants include,by way of non-limiting example, vegetable oils such as peanut oil,cottonseed oil, sesame oil, olive oil, corn oil, and oil of theobroma,glycerin, magnesium stearate, calcium stearate, stearic acid andcombinations thereof. In some embodiments, disintegrants are used tofacilitate disintegration of the tablet, and include, by way ofnon-limiting example, starches, clays, celluloses, algins, gums,crosslinked polymers and combinations thereof. Fillers include, by wayof non-limiting example, materials such as silicon dioxide, titaniumdioxide, alumina, talc, kaolin, powdered cellulose and microcrystallinecellulose, as well as soluble materials such as mannitol, urea, sucrose,lactose, dextrose, sodium chloride and sorbitol. In certain embodiments,stabilizers are used to inhibit or retard drug decomposition reactionsthat include, by way of example, oxidative reactions. In certainembodiments, surfactants are anionic, cationic, amphoteric or nonionicsurface active agents.

In some embodiments, ASBTIs, TGR5 agonists, or enteroendocrine peptidesecretion enhancing agents described herein are orally administered inassociation with a carrier suitable for delivery of the enteroendocrinepeptide secretion enhancing agents to the distal gastrointestinal tract(e.g., distal ileum, colon, and/or rectum).

In certain embodiments, a composition described herein comprises anASBTI, a TGR5 agonist, or an enteroendocrine peptide secretion enhancingagent in association with a matrix (e.g., a matrix comprisinghypermellose) that allows for controlled release of an active agent inthe distal part of the ileum and/or the colon. In some embodiments, acomposition comprises a polymer that is pH sensitive (e.g., a MMX™matrix from Cosmo Pharmaceuticals) and allows for controlled release ofan active agent in the distal part of the ileum. Examples of such pHsensitive polymers suitable for controlled release include and are notlimited to polyacrylic polymers (e.g., anionic polymers of methacrylicacid and/or methacrylic acid esters, e.g., Carbopol® polymers) thatcomprise acidic groups (e.g., —COOH, —SO₃H) and swell in basic pH of theintestine (e.g., pH of abut 7 to about 8). In some embodiments, acomposition suitable for controlled release in the distal ileumcomprises microparticulate active agent (e.g., micronized active agent).In some embodiments, a non-enzymatically degradingpoly(dl-lactide-co-glycolide) (PLGA) core is suitable for delivery of anenteroendocrine peptide secretion enhancing agent (e.g., bile acid) tothe distal ileum. In some embodiments, a dosage form comprising anenteroendocrine peptide secretion enhancing agent (e.g., bile acid) iscoated with an enteric polymer (e.g., Eudragit® S-100, cellulose acetatephthalate, polyvinylacetate phthalate, hydroxypropylmethylcellulosephthalate, anionic polymers of methacrylic acid, methacrylic acid estersor the like) for site specific delivery to the distal ileum and/or thecolon. In some embodiments, bacterially activated systems are suitablefor targeted delivery to the distal part of the ileum. Examples ofmicro-flora activated systems include dosage forms comprising pectin,galactomannan, and/or Azo hydrogels and/or glycoside conjugates (e.g.,conjugates of D-galactoside, β-D-xylopyranoside or the like) of theactive agent. Examples of gastrointestinal micro-flora enzymes includebacterial glycosidases such as, for example, D-galactosidase,β-D-glucosidase, α-L-arabinofuranosidase, β-D-xylopyranosidase or thelike.

The pharmaceutical composition described herein optionally include anadditional therapeutic compound described herein and one or morepharmaceutically acceptable additives such as a compatible carrier,binder, filling agent, suspending agent, flavoring agent, sweeteningagent, disintegrating agent, dispersing agent, surfactant, lubricant,colorant, diluent, solubilizer, moistening agent, plasticizer,stabilizer, penetration enhancer, wetting agent, anti-foaming agent,antioxidant, preservative, or one or more combination thereof. In someaspects, using standard coating procedures, such as those described inRemington's Pharmaceutical Sciences, 20th Edition (2000), a film coatingis provided around the formulation of the compound of Formula I. In oneembodiment, a compound described herein is in the form of a particle andsome or all of the particles of the compound are coated. In certainembodiments, some or all of the particles of a compound described hereinare microencapsulated. In some embodiments, the particles of thecompound described herein are not microencapsulated and are uncoated.

In certain embodiments, an oral formulation for use in any methoddescribed herein is, e.g., an ASBTI or an enteroendocrine peptidesecretion enhancing agent in association with a labile bile acidsequestrant. A labile bile acid sequestrant is a bile acid sequestrantwith a labile affinity for bile acids. In certain embodiments, a bileacid sequestrant described herein is an agent that sequesters (e.g.,absorbs or is charged with) bile acid, and/or the salts thereof.

In specific embodiments, the labile bile acid sequestrant is an agentthat sequesters (e.g., absorbs or is charged with) bile acid, and/or thesalts thereof, and releases at least a portion of the absorbed orcharged bile acid, and/or salts thereof in the distal gastrointestinaltract (e.g., the colon, ascending colon, sigmoid colon, distal colon,rectum, or any combination thereof). In certain embodiments, the labilebile acid sequestrant is an enzyme dependent bile acid sequestrant. Inspecific embodiments, the enzyme is a bacterial enzyme. In someembodiments, the enzyme is a bacterial enzyme found in highconcentration in human colon or rectum relative to the concentrationfound in the small intestine. Examples of micro-flora activated systemsinclude dosage forms comprising pectin, galactomannan, and/or Azohydrogels and/or glycoside conjugates (e.g., conjugates ofD-galactoside, β-D-xylopyranoside or the like) of the active agent.Examples of gastrointestinal micro-flora enzymes include bacterialglycosidases such as, for example, D-galactosidase, β-D-glucosidase,α-L-arabinofuranosidase, β-D-xylopyranosidase or the like. In someembodiments, the labile bile acid sequestrant is a time dependent bileacid sequestrant (i.e., the bile acid sequesters the bile acid and/orsalts thereof and after a time releases at least a portion of the bileacid and/or salts thereof). In some embodiments, a time dependent bileacid sequestrant is an agent that degrades in an aqueous environmentover time. In certain embodiments, a labile bile acid sequestrantdescribed herein is a bile acid sequestrant that has a low affinity forbile acid and/or salts thereof, thereby allowing the bile acidsequestrant to continue to sequester bile acid and/or salts thereof inan environ where the bile acids and/or salts thereof are present in highconcentration and release them in an environ wherein bile acids and/orsalts thereof are present in a lower relative concentration. In someembodiments, the labile bile acid sequestrant has a high affinity for aprimary bile acid and a low affinity for a secondary bile acid, allowingthe bile acid sequestrant to sequester a primary bile acid or saltthereof and subsequently release a secondary bile acid or salt thereofas the primary bile acid or salt thereof is converted (e.g.,metabolized) to the secondary bile acid or salt thereof. In someembodiments, the labile bile acid sequestrant is a pH dependent bileacid sequestrant. In some embodiments, the pH dependent bile acidsequestrant has a high affinity for bile acid at a pH of 6 or below anda low affinity for bile acid at a pH above 6. In certain embodiments,the pH dependent bile acid sequestrant degrades at a pH above 6.

In some embodiments, labile bile acid sequestrants described hereininclude any compound, e.g., a macro-structured compound, that cansequester bile acids and/or salts thereof through any suitablemechanism. For example, in certain embodiments, bile acid sequestrantssequester bile acids and/or salts thereof through ionic interactions,polar interactions, static interactions, hydrophobic interactions,lipophilic interactions, hydrophilic interactions, steric interactions,or the like. In certain embodiments, macrostructured compounds sequesterbile acids and/or sequestrants by trapping the bile acids and/or saltsthereof in pockets of the macrostructured compounds and, optionally,other interactions, such as those described above. In some embodiments,bile acid sequestrants (e.g., labile bile acid sequestrants) include, byway of non-limiting example, lignin, modified lignin, polymers,polycationic polymers and copolymers, polymers and/or copolymerscomprising anyone one or more of N-alkenyl-N-alkylamine residues; one ormore N,N,N-trialkyl-N—(N-alkenylamino)alkyl-azanium residues; one ormore N,N,N-trialkyl-N-alkenyl-azanium residues; one or morealkenyl-amine residues; or a combination thereof, or any combinationthereof.

Covalent Linkage of the Drug with a Carrier

In some embodiments, strategies used for colon targeted deliveryinclude, by way of non-limiting example, covalent linkage of the ASBTIand/or the enteroendocrine peptide secretion enhancing agents to acarrier, coating the dosage form with a pH-sensitive polymer fordelivery upon reaching the pH environment of the colon, using redoxsensitive polymers, using a time released formulation, utilizingcoatings that are specifically degraded by colonic bacteria, usingbioadhesive system and using osmotically controlled drug deliverysystems.

In certain embodiments of such oral administration of a compositioncontaining an ASBTI and or an enteroendocrine peptide secretionenhancing agent described herein involves covalent linking to a carrierwherein upon oral administration the linked moiety remains intact in thestomach and small intestine. Upon entering the colon the covalentlinkage is broken by the change in pH, enzymes, and/or degradation byintestinal microflora. In certain embodiments, the covalent linkagebetween the ASBTI and/or enteroendocrine peptide secretion enhancingagent and the carrier includes, by way of non-limiting example, azolinkage, glycoside conjugates, glucuronide conjugates, cyclodextrinconjugates, dextran conjugates, and amino-acid conjugates (highhydrophilicity and long chain length of the carrier amino acid).

Coating with Polymers: pH-Sensitive Polymers

In some embodiments, the oral dosage forms described herein are coatedwith an enteric coating to facilitate the delivery of an ASBTI and/or anenteroendocrine peptide secretion enhancing agent to the colon and/orrectum. In certain embodiments, an enteric coating is one that remainsintact in the low pH environment of the stomach, but readily dissolvedwhen the optimum dissolution pH of the particular coating is reachedwhich depends upon the chemical composition of the enteric coating. Thethickness of the coating will depend upon the solubility characteristicsof the coating material. In certain embodiments, the coating thicknessesused in such formulations described herein range from about 25 μm toabout 200 μm.

In certain embodiments, the compositions or formulations describedherein are coated such that an enteroendocrine peptide secretionenhancing agent of the composition or formulation is delivered to thecolon and/or rectum without absorbing at the upper part of theintestine. In a specific embodiment, specific delivery to the colonand/or rectum is achieved by coating of the dosage form with polymersthat degrade only in the pH environment of the colon. In alternativeembodiments, the composition is coated with an enteric coat thatdissolves in the pH of the intestines and an outer layer matrix thatslowly erodes in the intestine. In some of such embodiments, the matrixslowly erodes until only a core composition comprising anenteroendocrine peptide secretion enhancing agent (and, in someembodiments, an absorption inhibitor of the agent) is left and the coreis delivered to the colon and/or rectum.

In certain embodiments, pH-dependent systems exploit the progressivelyincreasing pH along the human gastrointestinal tract (GIT) from thestomach (pH 1-2 which increases to 4 during digestion), small intestine(pH 6-7) at the site of digestion and it to 7-8 in the distal ileum. Incertain embodiments, dosage forms for oral administration of thecompositions described herein are coated with pH-sensitive polymer(s) toprovide delayed release and protect the enteroendocrine peptidesecretion enhancing agents from gastric fluid. In certain embodiments,such polymers are be able to withstand the lower pH values of thestomach and of the proximal part of the small intestine, butdisintegrate at the neutral or slightly alkaline pH of the terminalileum and/or ileocecal junction. Thus, in certain embodiments, providedherein is an oral dosage form comprising a coating, the coatingcomprising a pH-sensitive polymer. In some embodiments, the polymersused for colon and/or rectum targeting include, by way of non-limitingexample, methacrylic acid copolymers, methacrylic acid and methylmethacrylate copolymers, Eudragit L100, Eudragit S100, Eudragit L-30D,Eudragit FS-30D, Eudragit L100-55, polyvinylacetate phthalate,hydroxypropyl ethyl cellulose phthalate, hydroxypropyl methyl cellulosephthalate 50, hydroxypropyl methyl cellulose phthalate 55, celluloseacetate trimelliate, cellulose acetate phthalate and combinationsthereof.

In certain embodiments, oral dosage forms suitable for delivery to thecolon and/or rectum comprise a coating that has a biodegradable and/orbacteria degradable polymer or polymers that are degraded by themicroflora (bacteria) in the colon. In such biodegradable systemssuitable polymers include, by way of non-limiting example, azo polymers,linear-type-segmented polyurethanes containing azo groups,polygalactomannans, pectin, glutaraldehyde crosslinked dextran,polysaccharides, amylose, guar gum, pectin, chitosan, inulin,cyclodextrins, chondroitin sulphate, dextrans, locust bean gum,chondroitin sulphate, chitosan, poly (-caprolactone), polylactic acidand poly(lactic-co-glycolic acid).

In certain embodiments of such oral administration of compositionscontaining one or more ASBTIs and/or enteroendocrine peptide secretionenhancing agents described herein, the compositions are delivered to thecolon without absorbing at the upper part of the intestine by coating ofthe dosage forms with redox sensitive polymers that are degraded by themicroflora (bacteria) in the colon. In such biodegradable systems suchpolymers include, by way of non-limiting example, redox-sensitivepolymers containing an azo and/or a disulfide linkage in the backbone.

In some embodiments, compositions formulated for delivery to the colonand/or rectum are formulated for time-release. In some embodiments, timerelease formulations resist the acidic environment of the stomach,thereby delaying the release of the enteroendocrine peptide secretionenhancing agents until the dosage form enters the colon and/or rectum.

In certain embodiments the time released formulations described hereincomprise a capsule (comprising an enteroendocrine peptide secretionenhancing agent and an optional absorption inhibitor) with hydrogelplug. In certain embodiments, the capsule and hydrogel plug are coveredby a water-soluble cap and the whole unit is coated with an entericpolymer. When the capsule enters the small intestine the enteric coatingdissolves and the hydrogels plug swells and dislodges from the capsuleafter a period of time and the composition is released from the capsule.The amount of hydrogel is used to adjust the period of time to therelease the contents.

In some embodiments, provided herein is an oral dosage form comprising amulti-layered coat, wherein the coat comprises different layers ofpolymers having different pH-sensitivities. As the coated dosage formmoves along GIT the different layers dissolve depending on the pHencountered. Polymers used in such formulations include, by way ofnon-limiting example, polymethacrylates with appropriate pH dissolutioncharacteristics, Eudragit® RL and Eudragit®RS (inner layer), andEudragit® FS (outer layer). In other embodiments the dosage form is anenteric coated tablets having an outer shell of hydroxypropylcelluloseor hydroxypropylmethylcellulose acetate succinate (HPMCAS).

In some embodiments, provided herein is an oral dosage form thatcomprises coat with cellulose butyrate phthalate, cellulose hydrogenphthalate, cellulose proprionate phthalate, polyvinyl acetate phthalate,cellulose acetate phthalate, cellulose acetate trimellitate,hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcelluloseacetate, dioxypropyl methylcellulose succinate, carboxymethylethylcellulose, hydroxypropyl methylcellulose acetate succinate,polymers and copolymers formed from acrylic acid, methacrylic acid, andcombinations thereof.

Combination Therapy

In certain instances, provided herein are combination compositionsand/or therapies comprising any compound described herein and anadditional therapeutic agent. In some embodiments, the additionaltherapeutic agent is a L-cell endocrine peptide enhancer. In someinstances, the L-cell endocrine peptide enhancer is a GLP-1 enhancer. Insome embodiments, the GLP-1 enhancer is GLP-1, a GLP-1 secretionenhancer, a GLP-1 degradation inhibitor, the like, or a combinationthereof. In certain instances, enhanced GLP-1 concentration provides areduction in food intake and/or a reduction in gastric emptying in humansubjects.

In some embodiments, the L-cell endocrine peptide enhancer is a GLP-2enhancer. In certain instances, the GLP-2 enhancer is GLP-2, a GLP-2secretion enhancer, a GLP-2 degradation inhibitor, the like, or acombination thereof. In certain instances, enhanced GLP-2 secretioninhibits gastric emptying and reduces intestinal permeability. In someinstances, enhanced GLP-2 secretion inhibits gastric acid secretion. Insome instances, enhanced GLP-2 secretion reduces or preventsinflammation in the gastrointestinal tract (gastrointestinal enteritis).In some instances, enhanced GLP-2 secretion regenerates and/or healsinjury to gastrointestinal tissues (e.g., radiation enteritis).

In some instances, the L-cell endocrine peptide enhancer is a PYYenhancer. In some instances, enhanced secretion of PYY provides areduction in sensation of hunger. In some instances, the L-cellendocrine peptide enhancer is an oxyntomodulin enhancer. In someinstances, the enhanced secretion of oxyntomodulin inhibitsmeal-stimulated gastric secretion.

TGR5 Receptor Modulators

In some instances, the additional therapeutic agent modulates bile acidreceptors in the gastrointestinal lumen. In some embodiments, theadditional therapeutic agent agonizes or partially agonizes bile acidreceptors (e.g., TGR5 receptors or Farnesoid-X receptors) in thegastrointestinal tract. In some embodiments, the additional therapeuticagent is a bile acid analog. In certain instances the additionaltherapeutic agent is a TGR5 agonist. In certain instances,administration of a TGR5 agonist in combination with any of thecompounds described herein enhances the secretion of enteroendocrinepeptides from L-cells. TGR5 modulators (e.g., agonists) include, and arenot limited to, the compounds described in, WO 2008/091540, WO2008/067219 and U.S. Appl. No. 2008/0221161.

Biguanides

In some embodiments, the additional therapeutic agent is a biguanide. Insome instances, biguanides reduce blood and/or plasma glucose levels.Examples of biguanides include and are not limited to metformin,buformin, phenformin, proguanil or the like.

Incretin Mimetics

In some embodiments, the additional therapeutic agent is an incretinmimetic. In some embodiments, an incretic mimic augments pancreasresponse to ingestion of food, In some instances, administration of anincretin mimetic in combination with any of the compounds describedherein lowers blood and/or plasma glucose levels. Examples of incretinmimetics include and are not limited to exenatide (Byetta®).

One currently used therapy for the treatment of diabetes is asubcutaneous injection of exenatide (Byetta®). In some embodiments, anoral combination of an ASBTI and a DPP-IV inhibitor is equally or moreeffective than an injection of exenatide in reducing plasma glucoselevels. In some embodiments, an oral combination of an ASBTI and aDPP-IV inhibitor reduces or eliminates discomfort associated withinjections of glucose-lowering medications.

Thiazolidinediones

In some embodiments, the additional therapeutic agent is athiazolidinedione. In some instances thiazolidinediones reverse insulinresistance and lower blood and/or plasma glucose levels. Examples ofthiazolidinediones include and are not limited to Rosiglitazone(Avandia), Pioglitazone (Actos), Troglitazone (Rezulin), MCC-555,rivoglitazone, ciglitazone or the like.

Enteroendocrine Peptides

In some embodiments, the additional therapeutic agent is anenteroendocrine peptide. In some embodiments, enteroendocrine peptidesreverse insulin resistance and lower blood and/or plasma glucose levels.Examples of enteroendocrine peptides that are administered as additionaltherapeutic agents include and are not limited to GLP-1 or GLP-1 analogssuch as Taspoglutide® (Ipsen), or the like.

Combination Therapy with ASBTI and DPP-IV Inhibitor

In specific embodiments, the additional therapeutic agent inhibitsdegradation of L-cell enteroendocrine peptides. In certain embodiments,the additional therapeutic agent is a DPP-IV inhibitor. In certaininstances, administration of an ASBTI to an individual in need thereofenhances the secretion of GLP-1; administration of a DPP-IV inhibitor incombination with the ASBTI reduces or inhibits degradation of GLP-1thereby prolonging the therapeutic benefit of enhanced levels of GLP-1.In some embodiments, administration of an ASBTI reduces weight of anindividual. In some embodiments, administration of an ASBTI incombination with a DPP-IV inhibitor reduces weight of an individual.

Another therapy that is current standard of care for the treatment ofdiabetes is a combination of metformin and sitagliptin (Janumet®). Atdoses of 0, 3, 30, 100, 300 mg/kg doses of metformin in combination with30 mg/kg of sitagliption, reductions in plasma glucose concentrationsare observed from 3 hours till about 6 hours post-dose (FIG. 3). In someembodiments, a combination of an ASBTI and sitagliptin maintains reducedplasma glucose concentrations for a longer duration of time (e.g., atleast 24 hours) compared to a combination of metformin and sitagliptin(about 6 hours) (FIGS. 1 and 2). In some instances ASBTI therapyeliminates side effects associated with metformin therapy and/or DPP-IVinhibitor therapy.

DPP-IV inhibitors suitable for use with the methods described hereininclude and are not limited to(2S)-1-{2-[(3-hydroxy-1-adamantyl)amino]acetyl}pyrrolidine-2-carbonitrile(vildagliptin),(3R)-3-amino-1-[9-(trifluoromethyl)-1,4,7,8-tetrazabicyclo[4.3.0]nona-6,8-dien-4-yl]-4-(2,4,5-trifluorophenyl)butan-1-one(sitagliptin),(1S,3S,5S)-2-[(2S)-2-amino-2-(3-hydroxy-1-adamantyl)acetyl]-2-azabicyclo[3.1.0]hexane-3-carbonitrile(saxagliptin), and2-({6-[(3R)-3-aminopiperidin-1-yl]-3-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl}methyl)benzonitrile(alogliptin).

In some embodiments of any of the methods described herein,administration of an ASBT inhibitor described herein in combination witha DPP-IV inhibitor increases the level of GLP-1 in the blood and/orplasma of an individual by from about 1.1 times to about 30 timescompared to the level of GLP-1 in the blood and/or plasma of theindividual prior to administration of the ASBTI in combination with theDPP-IV inhibitor. In some embodiments of any of the methods describedherein, administration of an ASBT inhibitor described herein incombination with a DPP-IV inhibitor increases the level of GLP-1 in theblood and/or plasma of an individual by from about 1.1 times to about 20times compared to the level of GLP-1 in the blood and/or plasma of theindividual prior to administration of the ASBTI in combination with theDPP-IV inhibitor. In some embodiments of any of the methods describedherein, administration of an ASBT inhibitor described herein incombination with a DPP-IV inhibitor increases the level of GLP-1 in theblood and/or plasma of an individual by from about 1.5 times to about 10times compared to the level of GLP-1 in the blood and/or plasma of theindividual prior to administration of the ASBTI in combination with theDPP-IV inhibitor. In some embodiments of any of the methods describedherein, administration of an ASBT inhibitor described herein incombination with a DPP-IV inhibitor increases the level of GLP-1 in theblood and/or plasma of an individual by from about 2 times to about 8times compared to the level of GLP-1 in the blood and/or plasma of theindividual prior to administration of the ASBTI in combination with theDPP-IV inhibitor. In some embodiments of any of the methods describedherein, administration of an ASBT inhibitor described herein incombination with a DPP-IV inhibitor increases the level of GLP-1 in theblood and/or plasma of an individual by from about 2 times to about 6times compared to the level of GLP-1 in the blood and/or plasma of theindividual prior to administration of the ASBTI in combination with theDPP-IV inhibitor. In some instances, an increase in GLP-1 level of fromabout 2 times to about 3 times following the administration of an ASBTinhibitor described herein in combination with a DPP-IV inhibitorcompared to the level of GLP-1 in the blood and/or plasma of theindividual prior to administration of the ASBTI in combination with theDPP-IV inhibitor is associated with an anti-diabetic effect. In someinstances, an increase in GLP-1 level of from about 3 times to about 8times following the administration of an ASBT inhibitor described hereinin combination with a DPP-IV inhibitor compared to the level of GLP-1 inthe blood and/or plasma of the individual prior to administration of theASBTI in combination with a DPP-IV inhibitor is associated withreduction in food intake and/or induction of satiety and/or weight loss.

In certain embodiments of any of the methods described herein,administration of an ASBTI in combination with a DPP-IV inhibitorreduces blood and/or plasma sugar levels by at least 20%, at least 30%,at least 40%, at least 50% at least 60%, at least 70% or at least 80%compared to blood and/or plasma sugar levels prior to administration ofthe ASBTI in combination with a DPP-IV inhibitor. In some embodiments ofany of the methods described herein, administration of an ASBTI incombination with a DPP-IV inhibitor reduces blood and/or plasma sugarlevels by at least 20% compared to blood and/or plasma sugar levelsprior to administration of the ASBTI in combination with a DPP-IVinhibitor. In some embodiments of any of the methods described herein,administration of an ASBTI in combination with a DPP-IV inhibitorreduces blood and/or plasma sugar levels by at least 30% compared toblood and/or plasma sugar levels prior to administration of the ASBTI incombination with a DPP-IV inhibitor. In some embodiments of any of themethods described herein, administration of an ASBTI in combination witha DPP-IV inhibitor reduces blood and/or plasma sugar levels by at least40% compared to blood and/or plasma sugar levels prior to administrationof the ASBTI in combination with a DPP-IV inhibitor.

In some embodiments of any of the methods described herein,administration of an ASBTI in combination with a DPP-IV inhibitorreduces blood and/or plasma sugar levels for a longer period of time(e.g., at least 24 hours) compared to reduction in blood and/or plasmasugar levels upon administration of metformin in combination with aDPP-IV inhibitor. In some embodiments of any of the methods describedherein, administration of a single dose of an ASBTI in combination witha DPP-IV inhibitor sustains reduced blood and/or plasma sugar levels forat least 6 hours, at least 12 hours, at least 14 hours, at least 16hours, at least 18 hours, at least 20 hours, at least 24 hours, at least30 hours, at least 36 hours or at least 48 hours compared to reductionin blood and/or plasma sugar levels upon administration of a single doseof metformin in combination with a DPP-IV inhibitor.

In some embodiments of any of the methods described herein,administration of an ASBTI in combination with a DPP-IV inhibitorresults in lower levels of intraenterocyte bile acids in an individualcompared to levels of intraenterocyte bile acids in an individualsuffering from necrotizing enterocolitis. In some embodiments of any ofthe methods described herein, administration of an ASBTI in combinationwith a DPP-IV inhibitor results in lower levels of necrosis and/ordamage to ileal architecture in an individual compared to levels ofnecrosis and/or damage to ileal architecture in an individual sufferingfrom necrotizing enterocolitis. In some embodiments of any of themethods described herein, administration of an ASBTI in combination witha DPP-IV inhibitor results in higher levels of GLP-1 in blood and/orplasma of an individual compared to levels of GLP-1 in blood and/orplasma of a normal individual. In some embodiments of any of the methodsdescribed herein, administration of an ASBTI in combination with aDPP-IV inhibitor results in higher levels of GLP-1 in blood and/orplasma of an individual compared to levels of GLP-1 in blood and/orplasma of an individual undergoing therapy with metformin and/or aDPP-IV inhibitor.

Combination Therapy with ASBTI, Biliary Shunt and DPP-IV Inhibitor

In some embodiments, an ASBTI is administered in combination with aDPP-IV inhibitor and/or a biliary shunt. Examples of biliary shuntsinclude and are not limited to the shunts described in WO 2007/0050628,the disclosure of biliary shunts described therein is incorporatedherein by reference. In some of such embodiments, a biliary shunt movesbile acid to the distal ileum and/or the rectum and/or the colon therebyincreasing the concentration of bile acids in the vicinity of L-cellspresent in the distal portion of the gastrointestinal tract. In someinstances such an increase in the concentration of bile acids in thevicinity of L-cells increases the secretion of GLP-1 from L-cellsthereby inducing satiey and/or reduction in hunger and/or weight lossand/or reduction in plasma glucose levels or any combination thereof.

An ASBTI and a second active ingredient are used such that thecombination is present in a therapeutically effective amount. Thattherapeutically effective amount arises from the use of a combination ofan ASBTI and the other active ingredient (e.g., a DPP-IV inhibitor)wherein each is used in a therapeutically effective amount, or by virtueof additive or synergistic effects arising from the combined use, eachcan also be used in a subclinical therapeutically effective amount,i.e., an amount that, if used alone, provides for reduced effectivenessfor the therapeutic purposes noted herein, provided that the combineduse is therapeutically effective. In some embodiments, the use of acombination of an ASBTI and any other active ingredient as describedherein encompasses combinations where the ASBTI or the other activeingredient is present in a therapeutically effective amount, and theother is present in a subclinical therapeutically effective amount,provided that the combined use is therapeutically effective owing totheir additive or synergistic effects. As used herein, the term“additive effect” describes the combined effect of two (or more)pharmaceutically active agents that is equal to the sum of the effect ofeach agent given alone. A syngergistic effect is one in which thecombined effect of two (or more) pharmaceutically active agents isgreater than the sum of the effect of each agent given alone. Anysuitable combination of an ASBIT with one or more of the aforementionedother active ingredients and optionally with one or more otherpharmacologically active substances is contemplated as being within thescope of the methods described herein.

In some embodiments, the particular choice of compounds depends upon thediagnosis of the attending physicians and their judgment of thecondition of the individual and the appropriate treatment protocol. Thecompounds are optionally administered concurrently (e.g.,simultaneously, essentially simultaneously or within the same treatmentprotocol) or sequentially, depending upon the nature of the disease,disorder, or condition, the condition of the individual, and the actualchoice of compounds used. In certain instances, the determination of theorder of administration, and the number of repetitions of administrationof each therapeutic agent during a treatment protocol, is based on anevaluation of the disease being treated and the condition of theindividual.

In some embodiments, therapeutically-effective dosages vary when thedrugs are used in treatment combinations. Methods for experimentallydetermining therapeutically-effective dosages of drugs and other agentsfor use in combination treatment regimens are described in theliterature.

In some embodiments of the combination therapies described herein,dosages of the co-administered compounds vary depending on the type ofco-drug employed, on the specific drug employed, on the disease orcondition being treated and so forth. In addition, when co-administeredwith one or more biologically active agents, the compound providedherein is optionally administered either simultaneously with thebiologically active agent(s), or sequentially. In certain instances, ifadministered sequentially, the attending physician will decide on theappropriate sequence of therapeutic compound described herein incombination with the additional therapeutic agent.

The multiple therapeutic agents (at least one of which is a therapeuticcompound described herein) are optionally administered in any order oreven simultaneously. If simultaneously, the multiple therapeutic agentsare optionally provided in a single, unified form, or in multiple forms(by way of example only, either as a single pill or as two separatepills). In certain instances, one of the therapeutic agents isoptionally given in multiple doses. In other instances, both areoptionally given as multiple doses. If not simultaneous, the timingbetween the multiple doses is any suitable timing, e.g, from more thanzero weeks to less than four weeks. In addition, the combinationmethods, compositions and formulations are not to be limited to the useof only two agents; the use of multiple therapeutic combinations arealso envisioned (including two or more compounds described herein).

In certain embodiments, a dosage regimen to treat, prevent, orameliorate the condition(s) for which relief is sought, is modified inaccordance with a variety of factors. These factors include the disorderfrom which the subject suffers, as well as the age, weight, sex, diet,and medical condition of the subject. Thus, in various embodiments, thedosage regimen actually employed varies and deviates from the dosageregimens set forth herein.

In some embodiments, the pharmaceutical agents which make up thecombination therapy described herein are provided in a combined dosageform or in separate dosage forms intended for substantially simultaneousadministration. In certain embodiments, the pharmaceutical agents thatmake up the combination therapy are administered sequentially, witheither therapeutic compound being administered by a regimen calling fortwo-step administration. In some embodiments, two-step administrationregimen calls for sequential administration of the active agents orspaced-apart administration of the separate active agents. In certainembodiments, the time period between the multiple administration stepsvaries, by way of non-limiting example, from a few minutes to severalhours, depending upon the properties of each pharmaceutical agent, suchas potency, solubility, bioavailability, plasma half-life and kineticprofile of the pharmaceutical agent.

In certain embodiments, ASBTI compounds described herein are combinedwith or utilized in combination with one or more of the followingtherapeutic agents in any combination: insulin, insulin-mimetics,incretin mimetics, GLP-1 or analogs thereof, GLP-2 or analogs thereof,oxyntomodulin, PYY, DPP-IV inhibitors, or TGR5 modulators.

In certain embodiments, provided herein are combination therapies. Incertain embodiments, the compositions described herein comprise anadditional therapeutic agent. In some embodiments, the methods describedherein comprise administration of a second dosage form comprising anadditional therapeutic agent. In certain embodiments, combinationtherapies the compositions described herein are administered as part ofa regimen. Therefore, additional therapeutic agents and/or additionalpharmaceutical dosage form can be applied to a patient either directlyor indirectly, and concomitantly or sequentially, with the compositionsand formulations described herein.

In certain embodiments the compositions described herein are used incombination with at least one appetite suppressant (e.g., a 5HTtransport inhibitor, a NE transport inhibitor, a CB-1 antagonist/inverseagonist, a ghrelin antagonist, a H3 antagonist/inverse agonist, a MCH1Rantagonist, a MCH2R agonist/antagonist, a NPY1 antagonist, a NPY2agonist, a mGluR5 antagonist, leptin, a leptin agonist/modulator, aleptin derivative, an opiod antagonist, an orexin antagonist, a BRS3agonist, a CCK-A agonist, CNTF, a CNTF agonist/modulator, a CNTFderivative, a 5HT2c agonist, a Mc5r agonist, a monoamine reuptakeinhibitor, a serotonin reuptake inhibitor, a GLP-1 agonist, axokine,fenfluramine, nalmafene, phentermine, rimonabant, sibutramine,topiramate, phytopharm compound 57, and combinations thereof). Incertain embodiments the compositions or formulations described hereinare used in combination with at least one metabolic rate enhancingagents (e.g., an ACC2 inhibitor, a β3 agonist, DGAT1 inhibitor, a DGAT2inhibitor, a FAS inhibitor, a PDE inhibitor, a thyroid hormone βagonist, an UCP-1, 2, or 3 activator, an acyl-estrogen, a glucocorticoidantagonist, an 11 β HSD-1 inhibitor, a Mc3r agonist, a SCD-1,oleoyl-estrone,3-[(3,5,7-trimethyl-1-adamantyl)methyl]-6,7,8,9-tetr-ahydro-5H-[1,2,4]triazolo[4,3-α]azepine;3-(1-adamantyl)-4-ethyl-5-(e-thylthio)-4H-1,2,4-triazole;3-adamantanyl-4,5,6,7,8,9,10,11,12,3a-de-cahydro-1,2,4-triazolo[4,3-α][11]annulene,3-(1-adamantyl)-5-(3,4,5-trimethoxyphenyl)-4-methyl-4H-1,2,4-triazoleand combinations thereof). In some embodiments, an enteroendocrinepeptide secretion enhancing agent is combined or administered with aphosphodiesterase inhibitor. In certain embodiments, an enteroendocrinepeptide secretion enhancing agent is combined or administered withcaffeine. In certain embodiments the compositions or formulationsdescribed herein are used in combination with at least one nutrientabsorption inhibitors (e.g., a lipase inhibitor; a fatty acidtransporter inhibitor; a dicarboxylate transporter inhibitor; a glucosetransporter inhibitor; a phosphate transporter inhibitor; orlistat andcombinations thereof). In certain embodiments the compositions orformulations described herein are used in combination with at least oneappetite suppressant and at least one metabolic rate enhancing agents.In certain embodiments the compositions or formulations described hereinare used in combination with at least one appetite suppressant and atleast one nutrient absorption inhibitors. In certain embodiments thecompositions or formulations described herein are used in combinationwith at least one nutrient absorption inhibitors and at least onemetabolic rate enhancing agents. In certain embodiments the compositionsor formulations described herein are used in combination with at leastone appetite suppressant, at least one metabolic rate enhancing agentsand at least one nutrient absorption inhibitors.

In some embodiments, enteroendocrine peptide secretion enhancing agentstherapies described herein are used in combination with biliary shunttreatments. In certain embodiments, a bilary shunt is a stentedthin-walled catheter placed endoscopically within the common bile duct.In certain embodiments, the catheter runs a length down thegastrointestinal tract, providing for delivery of bile acids and/orsalts to the distal gastrointestinal tract. FIG. 16 illustratesplacement of a biliary shunt useful for delivery of endogenous bileacids and/or salts to the distal gastrointestinal tract (e.g., distalileum, colon, and/or rectum).

In some embodiments, a combination described herein comprises a DPP-4(used interchangeably herein with DPP-IV) inhibitor. In certainembodiments, a method described herein comprises administering a DPP-4inhibitor. In some instances, inhibition of DPP-IV reduces thedegradation of enteroendocrine peptide products (e.g. GLP-1) therebyprolonging the effect of GLP-1 in reducing blood glucose levels.

DPP-IV inhibitors suitable for use with the methods described hereininclude and are not limited to(2S)-1-{2-[(3-hydroxy-1-adamantyl)amino]acetyl}pyrrolidine-2-carbonitrile(vildagliptin),(3R)-3-amino-1-[9-(trifluoromethyl)-1,4,7,8-tetrazabicyclo[4.3.0]nona-6,8-dien-4-yl]-4-(2,4,5-trifluorophenyl)butan-1-one(sitagliptin),(1S,3S,5S)-2-[(2S)-2-amino-2-(3-hydroxy-1-adamantyl)acetyl]-2-azabicyclo[3.1.0]hexane-3-carbonitrile(saxagliptin), and2-({6-[(3R)-3-aminopiperidin-1-yl]-3-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl}methyl)benzonitrile(alogliptin).

In some embodiments of any of the methods described herein,administration of an enteroendocrine peptide secretion enhancing agentsdescribed herein in combination with a DPP-IV inhibitor increases thelevel of GLP-1 in the blood and/or plasma of an individual by from about1.1 times to about 30 times compared to the level of GLP-1 in the bloodand/or plasma of the individual prior to administration of theenteroendocrine peptide secretion enhancing agent in combination withthe DPP-IV inhibitor. In some embodiments of any of the methodsdescribed herein, administration of an enteroendocrine peptide secretionenhancing agent described herein in combination with a DPP-IV inhibitorincreases the level of GLP-1 in the blood and/or plasma of an individualby from about 1.1 times to about 20 times compared to the level of GLP-1in the blood and/or plasma of the individual prior to administration ofthe enteroendocrine peptide secretion enhancing agent in combinationwith the DPP-IV inhibitor. In some embodiments of any of the methodsdescribed herein, administration of an enteroendocrine peptide secretionenhancing agent described herein in combination with a DPP-IV inhibitorincreases the level of GLP-1 in the blood and/or plasma of an individualby from about 1.5 times to about 10 times compared to the level of GLP-1in the blood and/or plasma of the individual prior to administration ofthe enteroendocrine peptide secretion enhancing agent in combinationwith the DPP-IV inhibitor. In some embodiments of any of the methodsdescribed herein, administration of an enteroendocrine peptide secretionenhancing agent described herein in combination with a DPP-IV inhibitorincreases the level of GLP-1 in the blood and/or plasma of an individualby from about 2 times to about 8 times compared to the level of GLP-1 inthe blood and/or plasma of the individual prior to administration of theenteroendocrine peptide secretion enhancing agent in combination withthe DPP-IV inhibitor. In some embodiments of any of the methodsdescribed herein, administration of an enteroendocrine peptide secretionenhancing agent described herein in combination with a DPP-IV inhibitorincreases the level of GLP-1 in the blood and/or plasma of an individualby from about 2 times to about 6 times compared to the level of GLP-1 inthe blood and/or plasma of the individual prior to administration of theenteroendocrine peptide secretion enhancing agent in combination withthe DPP-IV inhibitor. In some instances, an increase in GLP-1 level offrom about 2 times to about 3 times following the administration of anenteroendocrine peptide secretion enhancing agent inhibitor describedherein in combination with a DPP-IV inhibitor compared to the level ofGLP-1 in the blood and/or plasma of the individual prior toadministration of the enteroendocrine peptide secretion enhancing agentin combination with the DPP-IV inhibitor is associated with ananti-diabetic effect. In some instances, an increase in GLP-1 level offrom about 3 times to about 8 times following the administration of anenteroendocrine peptide secretion enhancing agent described herein incombination with a DPP-IV inhibitor compared to the level of GLP-1 inthe blood and/or plasma of the individual prior to administration of theenteroendocrine peptide secretion enhancing agent in combination with aDPP-IV inhibitor is associated with reduction in food intake and/orinduction of satiety and/or weight loss.

In certain embodiments of any of the methods described herein,administration of an enteroendocrine peptide secretion enhancing agentin combination with a DPP-IV inhibitor reduces blood and/or plasma sugarlevels by at least 20%, at least 30%, at least 40%, at least 50% atleast 60%, at least 70% or at least 80% compared to blood and/or plasmasugar levels prior to administration of the enteroendocrine peptidesecretion enhancing agent in combination with a DPP-IV inhibitor. Insome embodiments of any of the methods described herein, administrationof an enteroendocrine peptide secretion enhancing agent in combinationwith a DPP-IV inhibitor reduces blood and/or plasma sugar levels by atleast 20% compared to blood and/or plasma sugar levels prior toadministration of the enteroendocrine peptide secretion enhancing agentin combination with a DPP-IV inhibitor. In some embodiments of any ofthe methods described herein, administration of an enteroendocrinepeptide secretion enhancing agent in combination with a DPP-IV inhibitorreduces blood and/or plasma sugar levels by at least 30% compared toblood and/or plasma sugar levels prior to administration of theenteroendocrine peptide secretion enhancing agent in combination with aDPP-IV inhibitor. In some embodiments of any of the methods describedherein, administration of an enteroendocrine peptide secretion enhancingagent in combination with a DPP-IV inhibitor reduces blood and/or plasmasugar levels by at least 40% compared to blood and/or plasma sugarlevels prior to administration of the enteroendocrine peptide secretionenhancing agent in combination with a DPP-IV inhibitor.

In some embodiments of any of the methods described herein,administration of an enteroendocrine peptide secretion enhancing agentin combination with a DPP-IV inhibitor reduces blood and/or plasma sugarlevels for a longer period of time (e.g., at least 24 hours) compared toreduction in blood and/or plasma sugar levels upon administration ofmetformin in combination with a DPP-IV inhibitor. In some embodiments ofany of the methods described herein, administration of a single dose ofan enteroendocrine peptide secretion enhancing agent in combination witha DPP-IV inhibitor sustains reduced blood and/or plasma sugar levels forat least 6 hours, at least 12 hours, at least 14 hours, at least 16hours, at least 18 hours, at least 20 hours, at least 24 hours, at least30 hours, at least 36 hours or at least 48 hours compared to reductionin blood and/or plasma sugar levels upon administration of a single doseof metformin in combination with a DPP-IV inhibitor.

In some embodiments of any of the methods described herein,administration of an enteroendocrine peptide secretion enhancing agentin combination with a DPP-IV inhibitor results in higher levels of GLP-1in blood and/or plasma of an individual compared to levels of GLP-1 inblood and/or plasma of a normal individual. In some embodiments of anyof the methods described herein, administration of an enteroendocrinepeptide secretion enhancing agent in combination with a DPP-IV inhibitorresults in higher levels of GLP-1 in blood and/or plasma of anindividual compared to levels of GLP-1 in blood and/or plasma of anindividual undergoing therapy with metformin and/or a DPP-IV inhibitor.

Kits

In another aspect, provided herein are kits containing a device forrectal administration pre-filled a pharmaceutical composition describedherein. In certain embodiments, kits contain a device for rectaladministration and a pharmaceutical composition (e.g., a rectal dosageform) as described herein. In certain embodiments the kits includesprefilled bags for administration of rectal enemas, while in otherembodiments the kits include prefilled bags for administration of rectalgels. In certain embodiments the kits includes prefilled syringes foradministration of rectal enemas, while in other embodiments the kitsinclude prefilled syringes for administration of rectal gels. In certainembodiments the kits includes prefilled pressurized cans foradministration of rectal foams.

Pharmaceutical Compositions

Provided herein, in certain embodiments, is a pharmaceutical compositioncomprising a therapeutically effective amount of any compound describedherein. In certain instances, the pharmaceutical composition comprisesan ASBT inhibitor (e.g., any ASBTI described herein).

In certain embodiments, pharmaceutical compositions are formulated in aconventional manner using one or more physiologically acceptablecarriers including, e.g., excipients and auxiliaries which facilitateprocessing of the active compounds into preparations which are suitablefor pharmaceutical use. In certain embodiments, proper formulation isdependent upon the route of administration chosen. A summary ofpharmaceutical compositions described herein is found, for example, inRemington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton,Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington'sPharmaceutical Sciences, Mack Publishing Co., Easton, Pa. 1975;Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms,Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms andDrug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999).

A pharmaceutical composition, as used herein, refers to a mixture of acompound described herein, such as, for example, a compound of FormulaI-VI, with other chemical components, such as carriers, stabilizers,diluents, dispersing agents, suspending agents, thickening agents,and/or excipients. In certain instances, the pharmaceutical compositionfacilitates administration of the compound to an individual or cell. Incertain embodiments of practicing the methods of treatment or useprovided herein, therapeutically effective amounts of compoundsdescribed herein are administered in a pharmaceutical composition to anindividual having a disease, disorder, or condition to be treated. Inspecific embodiments, the individual is a human. As discussed herein,the compounds described herein are either utilized singly or incombination with one or more additional therapeutic agents.

In certain embodiments, the pharmaceutical formulations described hereinare administered to an individual in any manner, including one or moreof multiple administration routes, such as, by way of non-limitingexample, oral, parenteral (e.g., intravenous, subcutaneous,intramuscular), intranasal, buccal, topical, rectal, or transdermaladministration routes.

In certain embodiments, a pharmaceutical compositions described hereinincludes one or more compound described herein as an active ingredientin free-acid or free-base form, or in a pharmaceutically acceptable saltform. In some embodiments, the compounds described herein are utilizedas an N-oxide or in a crystalline or amorphous form (i.e., a polymorph).In some situations, a compound described herein exists as tautomers. Alltautomers are included within the scope of the compounds presentedherein. In certain embodiments, a compound described herein exists in anunsolvated or solvated form, wherein solvated forms comprise anypharmaceutically acceptable solvent, e.g., water, ethanol, and the like.The solvated forms of the compounds presented herein are also consideredto be described herein.

A “carrier” includes, in some embodiments, a pharmaceutically acceptableexcipient and is selected on the basis of compatibility with compoundsdescribed herein, such as, compounds of any of Formula I-VI, and therelease profile properties of the desired dosage form. Exemplary carriermaterials include, e.g., binders, suspending agents, disintegrationagents, filling agents, surfactants, solubilizers, stabilizers,lubricants, wetting agents, diluents, and the like. See, e.g.,Remington. The Science and Practice of Pharmacy, Nineteenth Ed (Easton,Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington'sPharmaceutical Sciences, Mack Publishing Co., Easton, Pa. 1975;Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms,Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms andDrug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999).

Moreover, in certain embodiments, the pharmaceutical compositionsdescribed herein are formulated as a dosage form. As such, in someembodiments, provided herein is a dosage form comprising a compounddescribed herein, suitable for administration to an individual. Incertain embodiments, suitable dosage forms include, by way ofnon-limiting example, aqueous oral dispersions, liquids, gels, syrups,elixirs, slurries, suspensions, solid oral dosage forms, aerosols,controlled release formulations, fast melt formulations, effervescentformulations, lyophilized formulations, tablets, powders, pills,dragees, capsules, delayed release formulations, extended releaseformulations, pulsatile release formulations, multiparticulateformulations, and mixed immediate release and controlled releaseformulations.

Release in Distal Ileum and/or Colon

In certain embodiments, a dosage form comprises a matrix (e.g., a matrixcomprising hypermellose) that allows for controlled release of an activeagent in the distal jejunum, proximal ileum, distal ileum and/or thecolon. In some embodiments, a dosage form comprises a polymer that is pHsensitive (e.g., a MMX™ matrix from Cosmo Pharmaceuticals) and allowsfor controlled release of an active agent in the ileum and/or the colon.Examples of such pH sensitive polymers suitable for controlled releaseinclude and are not limited to polyacrylic polymers (e.g., anionicpolymers of methacrylic acid and/or methacrylic acid esters, e.g.,Carbopol® polymers) that comprise acidic groups (e.g., —COOH, —SO₃H) andswell in basic pH of the intestine (e.g., pH of about 7 to about 8). Insome embodiments, a dosage form suitable for controlled release in thedistal ileum comprises microparticulate active agent (e.g., micronizedactive agent). In some embodiments, a non-enzymatically degradingpoly(dl-lactide-co-glycolide) (PLGA) core is suitable for delivery of anASBTI to the distal ileum. In some embodiments, a dosage form comprisingan ASBTI is coated with an enteric polymer (e.g., Eudragit® S-100,cellulose acetate phthalate, polyvinylacetate phthalate,hydroxypropylmethylceilulose phthalate, anionic polymers of methacrylicacid, methacrylic acid esters or the like) for site specific delivery tothe ileum and/or the colon. In some embodiments, bacterially activatedsystems are suitable for targeted delivery to the ileum. Examples ofmicro-flora activated systems include dosage forms comprising pectin,galactomannan, and/or Azo hydrogels and/or glycoside conjugates (e.g.,conjugates of D-galactoside, β-D-xylopyranoside or the like) of theactive agent. Examples of gastrointestinal micro-flora enzymes includebacterial glycosidases such as, for example, D-galactosidase,β-D-glucosidase, α-L-arabinofuranosidase, β-D-xylopyranosidase or thelike.

The pharmaceutical solid dosage forms described herein optionallyinclude an additional therapeutic compound described herein and one ormore pharmaceutically acceptable additives such as a compatible carrier,binder, filling agent, suspending agent, flavoring agent, sweeteningagent, disintegrating agent, dispersing agent, surfactant, lubricant,colorant, diluent, solubilizer, moistening agent, plasticizer,stabilizer, penetration enhancer, wetting agent, anti-foaming agent,antioxidant, preservative, or one or more combination thereof. In someaspects, using standard coating procedures, such as those described inRemington's Pharmaceutical Sciences, 20th Edition (2000), a film coatingis provided around the formulation of the compound of Formula I-VI. Inone embodiment, a compound described herein is in the form of a particleand some or all of the particles of the compound are coated. In certainembodiments, some or all of the particles of a compound described hereinare microencapsulated. In some embodiments, the particles of thecompound described herein are not microencapsulated and are uncoated.

An ASBT inhibitor (e.g., a compound of Formula I-VI) is used in thepreparation of medicaments for the prophylactic and/or therapeutictreatment of obesity and/or diabetes. A method for treating any of thediseases or conditions described herein in an individual in need of suchtreatment, involves administration of pharmaceutical compositionscontaining at least one ASBT inhibitor described herein, or apharmaceutically acceptable salt, pharmaceutically acceptable N-oxide,pharmaceutically active metabolite, pharmaceutically acceptable prodrug,or pharmaceutically acceptable solvate thereof, in therapeuticallyeffective amounts to said individual.

Screening Process

Provided in certain embodiments herein are processes and kits foridentifying compounds suitable for treating obesity, metabolic disordersor disorders mediated by L-cell enteroendocrine peptides. In certainembodiments, provided herein are assays for identifying compounds thatselectively inhibit the ASBT, or enhance the secretion of L-cellenteroendocrine peptides, or a combination thereof by:

-   -   a. providing cells that are a model of intestinal L-cells (e.g.,        SLC-1 cells, GLUTag cells, NCI-H719 cells);    -   b. contacting the cells with a compound (e.g., a compound as        described herein);    -   c. detecting or measuring the effect of the compound on the        secretion of enteroendocrine peptides (e.g., GLP-1, GLP-2) from        the cells.

In certain embodiments, provided herein are assays for identifyingcompounds that are non-systemic compounds by

-   -   a. providing cells that are a model of intestinal permeability        (e.g., Caco-2 cells);    -   b. culturing the cells as a monolayer on semi-permeable plastic        supports that are fitted into the wells of multi-well culture        plates;    -   c. contacting the apical or basolateral surface of the cells        with a compound (e.g., a compound as described herein) and        incubating for a suitable length of time;    -   d. detecting or measuring the concentration of the compound on        both sides of the monolayer by liquid-chromatography-mass        spectrometry (LC-MS) and computing intestinal permeability of        the compound.

In certain embodiments, non-systemic compounds are identified bysuitable parallel artificial membrane permeability assays (PAMPA).

In certain embodiments, non-systemic compounds are identified by use ofisolated vascular-perfused gut preparations.

In certain embodiments, provided herein are assays for identifyingcompounds that inhibit recycling of bile acid salts by

-   -   a. providing cells that are a model of intestinal cells with        apical bile acid tranporters (e.g., BHK cells, CHO cells);    -   b. incubating the cells with a compound (e.g., a compound as        described herein) and/or a radiolabeled bile acid (e.g., ¹⁴C        taurocholate) for a suitable length of time;    -   c. washing the cells with a suitable buffer (e.g. phosphate        buffered saline);    -   d. detecting or measuring the residual concentration of the        radiolabeled bile acid in the cells.

EXAMPLES Example 1 Synthesis of1-phenethyl-1-((1,4-diazabicyclo[2.2.2]octanyl)pentyl)imidodicarbonimidicdiamide, Iodide Salt

Step 1: Synthesis of 5-(1,4-diazabicyclo[2.2.2]octanyl)-1-iodo pentane,iodide salt

1,4-diazabicyclo[2.2.2]octane is suspended in THF. Diiodopentane isadded dropwise and the mixture is refluxed overnight. The reactionmixture is filtered.

Step 2: Synthesis ofN-phenethyl-5-(1,4-diazabicyclo[2.2.2]octanyl)-1-iodo pentane, IodideSalt

5-(1,4-diazabicyclo[2.2.2]octanyl)-1-iodo pentane, iodide salt issuspended in acetonitrile. Phenethylamine is added dropwise and themixture is refluxed overnight. The reaction mixture is filtered.

Step 3: Synthesis of1-phenethyl-1-((1,4-diazabicyclo[2.2.2]octanyl)pentyl)imidodicarbonimidicdiamide, iodide salt

N-phenethyl-5-(1,4-diazabicyclo[2.2.2]octanyl)-1-iodo pentane, iodidesalt is heated with dicyanodiamide in n-butanol for 4 h. The reactionmixture is concentrated under reduced pressure.

The compounds in Table I are prepared using methods as described herein,and using appropriate starting materials.

TABLE 1 Compound No. Structure 1

2

3

4

5

6

7

8

9

10

11

Example 2

In Vitro Assay for Inhibition of ASBT-Mediated Bile Acid Uptake

Baby hamster kidney (BHK) cells are transfected with cDNA of human ASBT.The cells are seeded in 96-well tissue culture plates at 60,000cells/well. Assays are run within 24 hours of seeding.

On the day of the assay the cell monolayer is washed with 100 mL ofassay buffer. The test compound is added to each well along with 6 mM[¹⁴C] taurocholate in assay buffer (final concentration of 3 mM [¹⁴C]taurocholate in each well). The cell cultures are incubated for 2 h at37° C. The wells are washed with PBS. Scintillation counting fluid isadded to each well, the cells are shaken for 30 minutes prior tomeasuring amount of radioactivity in each well. A test compound that hassignificant ASBT inhibitory activity provides an assay wherein lowlevels of radioactivity are observed in the cells.

Example 3

In Vitro Assay for Secretion of GLP-1

Human NCI-H716 cells are used as a model for L-cells. Two days beforeeach assay experiment, cells are seeded in 12-well culture plates coatedwith Matrigel® to induce cell adhesion. On the day of the assay, cellsare washed with buffer. The cells are incubated for 2 hours with mediumalone, or with test compound. The extracellular medium is assayed forthe presence of GLP-1. Peptides in the medium are collected by reversephase adsorption and the extracts are stored until assay. The presenceof GLP-1 is assayed using an antiserum directed against the carboxylterminus of GLP-1 amide (total immunoreactive GLP-1; Affinity ResearchProducts, Nottingham, UK). The detection of increased levels of GLP-1 ina well containing a test compound identifies the test compound as acompound that can enhance GLP-1 secretions from L-cells.

Example 4

In Vivo Bioavailability Assay

The test compounds are solubilized in saline solutions. Sprague Dawleyrats are dosed at 2-10 mg/kg body weight by iv and oral dosing.Peripheral blood samples are taken from the femoral artery at selectedtime periods up to 8 hours. Plasma concentrations of the compounds aredetermined by quantitative HPLC and/or mass spectrometry. Clearance andAUC values are determined for the compounds.

For oral dosing, bioavailabilty is calculated by also drawing plasmasamples from the portal vein. Cannulae are inserted in the femoralartery and the hepatic portal vein to obtain estimates of totalabsorption of drug without first-pass clearance in the liver. Thefraction absorbed (F) is calculated byF=AUC_(po)/AUC_(iv)

Example 5

In Vivo Animal Model of Necrotizing Enterocolitis and Obesity

Sprague-Dawley (SD) rats are used as a model of necrotizingenterocolitis (NEC). NEC is induced by collecting premature SD rats bycesarean section 1 day prior to scheduled birth. SD rat pups are fedwith cow's milk-based formula and subjected to asphyxia and cold stresstwice daily to develop NEC.

A non-genetic mouse obesity model is used. Obesity is induced in acohort of C₅₇BL/6 (wild-type) mice, by a high-fat cafeteria diet. Themice are administered 30 mg/kg of compound 8 once daily as an oralsolution in saline. The mice are weighed daily over a period of twoweeks. Cafeteria dieting is continued throughout the experiment. Obesemice administered a saline solution alone (placebo) serve as negativecontrols. The weight gain in the control group is compared with theweight gain or loss of the test group.

Example 6

Histological Evaluations of Necrosis or Damage to Ileal Architecture orIleal Cells in NEC

Histological sections are obtained after administration of an ASBTIand/or an enteroendocrine peptide enhancing agent and/or a FXR agonistto the ileum of NEC suffering SD rats and compared to the correspondinghistological sections of a normal SD rat. Histological changes arescored by a blind evaluator and graded using the following scoringsystem:

Score Histological observations 0 Normal, no necrosis or damage 1 Verymild, slight submucosal and/or lamina propria separation 2 Mild, somesubmucosal and/or lamina propria separation 3 Moderate, moderatesubmucosal and/or lamina propria separation 4 Severe, severe submucosaland/or lamina propria separation and/or severe edema in submucosa andmuscular layers, regional villous sloughing 5 Necrosis, loss of villiand loss of ileal architectureHistological scores greater than or equal to 3 are considered to havedeveloped NEC.

A single administration of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideis administered to produce a dose-dependent reduction of thehistological score. Control mice are treated with only the controlvehicle (saline in 10% of DMSO).

Example 6B

Assay to Determine Ileal Intraenterocyte and Luminal Bile Acid Levels

Ileal luminal bile acid levels in SD rats are determined by flusing a3-cm section of distal ileum with sterile, cold PBS. After flushing withadditional PBS, the same section of ileum is weighed and thenhomogenized in fresh PBS for determination of interenterocyte bile acidlevels. A LC/MS/MS system is used to evaluate cholic acid, DCA, LCA,chenodeoxycholic acid, and ursodeoxycholic acid levels.

Example 7A

In Vivo Assay for Reduction in Blood Glucose Levels

Male db/db mice 6-7 weeks (approx 30 g) were fasted for 2 h. Acombination of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxide(Compound 100A) at doses of 0, 0.1, 1, 10, mg/kg in 10% of DMSO-waterand 30 mg/kg sitagliptin (DPP-IV inhibitor) in a mixture ofvaline-pyrrolidine in water was administered orally to male db/db mice(n=7-10 per group). Blood sample (˜1 μl from tail for glucose leveltesting) was taken at t=−24 h, 0, 60, 180, 240, 300, 360 min and 24 hafter administration.

A single administration of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxidein combination with sitagliptin produced a dose-dependent reduction ofthe elevated blood glucose levels compared to the control mice treatedwith sitagliptin alone or with vehicle solution of 10% of DMSO (FIG. 1).Reductions in plasma glucose concentrations are observed from 3 hours upto 24 hours post-dose and accompanied after 6 hours by a 5.5%, 5.6%, 15%and 42.3% reduction in plasma glucose from baseline, (doses of 0, 01, 1and 10 mg/kg of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxiderespectively). Plasma glucose concentrations in the control mice treatedwith only the control vehicle (saline in 10% of DMSO) increased by 1.5%after 6 h. Administration of the ASBTI alone also lowers plasma glucoseconcentrations in male db/db mice as shown in FIG. 1. Weight loss isshown in FIG. 9.

Example 7B

In Vivo Assay for Reduction in Blood Glucose Levels

Male db/db mice 8-9 weeks (approx 40 g) were fasted for 2 h. Acombination of1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate (Compound 100B) at doses of 0, 3, 30, 100 mg/kg insaline-water and 30 mg/kg sitagliptin (DPP-IV inhibitor) in a mixture ofvaline-pyrrolidine in water was administered orally to male db/db mice(n=6-11 per group). Blood sample (˜1 μl from tail for glucose leveltesting) was taken at t=−24 h, 0, 1, 3, 4, 5, 6 h and 24 h afteradministration.

A single administration of1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate in combination with sitagliptin produced adose-dependent reduction of the elevated blood glucose levels comparedto the control mice treated with sitagliptin alone or with vehiclesolution of saline-water (FIG. 2). Reductions in plasma glucoseconcentrations are observed from 3 hours till 24 hours post-dose andaccompanied after 6 hours by a 5.5%, 5.6%, 15% and 42.3% reduction inplasma glucose from baseline, (doses of 0, 3, 30 and 100 mg/kg of1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate respectively). Plasma glucose concentrations in thecontrol mice treated with only the control vehicle (saline-water)increased by 1.5% after 6 h (FIG. 2). Weight loss is shown in. FIG. 8.

Administration of doses of1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate alone also lowers plasma glucose levels in dosedependent manner (FIG. 4).

Example 8

Investigation of orally delivered1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octaneMethane Sulfonate (Compound 100B) and Metformin in Combination withDPP-IV Inhibitor on Plasma GLP-1 Levels in Normal Rats

12-week-old male HSD rats were fasted for 16 h and given oral dose of 0,3, 30, 100 mg/kg of the ASBTI1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate (Synthesized by Nanosyn Inc., CA, USA) or metformin(Control, 0, 3, 30, 100, 300 mg/kg) in saline and a dose of 30 mg/kgsitaglipin in a mixture of valine-pyrrolidine in water (n=5 per group).Blood samples in volume of 0.6 ml for each time point were taken fromthe caudal vein with a heparinized capillary tube 0, 1, 3 and 5 h afterthe administration of compounds and plasma GLP-1 level were determined.Aprotinin and 10 μl of DPP-IV inhibitor per ml of blood were used forblood sample preservation during 10 min centrifugation and for storageat −70° C. or below. GLP-1 (Active pM) was tested by Millipore ELISAKits (Millipore Corporation, 290 Concord Road, Billerica, Mass.). ASBTIalone or ASBTI in combination with sitagliptin elicited an elevation inGLP-1 levels in a dose-dependent manner (FIG. 6, FIG. 7) in normal rats.FIG. 5 shows elevation in GLP-1 levels in plasma of normal rats uponadministration of a combination of metformin and sitagliptin.

Example 9

Investigation of Orally Delivered Compound 100A and 100B in Combinationwith DPP-IV Inhibitor Versus Subcutaneous Exenatide

A similar experiment as described above is carried out to test doses of0, 0.01, 1 and 10 mg/kg of the ASBTI(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxide(Compound 100A) in combination with 30 mg/kg sitagliptin and asubcutaneous injection of exenatide. An oral dose of a combination of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxide(10 mg/kg) in combination with 30 mg/kg. sitagliptin reduces plasmaglucose levels by 35% while a sub-cutaneous injection of exenatide (0.2mg/kg) reduces plasma glucose level by 31% at 3 hours post-dose (FIG.10).

A similar experiment as described above is carried out to compareglucose lowering effect of orally administered doses of 0, 3, 30, and100, mg/kg of the ABTI1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate (Compound 100B) in combination with a 30 mg/kg dose ofsitagliptin versus 0.02, 0.2 and 2 mg/kg doses of exenatide. FIG. 10shows that an oral combination of1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate in combination with a 30 mg/kg dose of sitagliptinreduces blood glucose levels more than a single subcutaneous injectionof 0.02 mg/kg exenatide at 3 hours after dosing.

Example 10

Tablet Formulation

10 kg of a compound of Formula I-VI is first screened through a suitablescreen (e.g. 500 micron). 25 kg Lactose monohydrate, 8 kghydroxypropylmethyl cellulose, the screened compound of Formula I-VI and5 kg calcium hydrogen phosphate (anhydrous) are then added to a suitableblender (e.g. a tumble mixer) and blended. The blend is screened througha suitable screen (e.g. 500 micron) and reblended. About 50% of thelubricant (2.5 kg, magnesium stearate) is screened, added to the blendand blended briefly. The remaining lubricant (2 kg, magnesium stearate)is screened, added to the blend and blended briefly. The granules arescreened (e.g. 200 micron) to obtain granulation particles of thedesired size. In some embodiments, the granules are optionally coatedwith a drug release controlling polymer such as polyvinylpyrrolidine,hydroxypropylcellulose, hydroxypropylmethyl cellulose, methyl cellulose,or a methacrylic acid copolymer, to provide an extended releaseformulation. The granules are filled in gelatin capsules.

Example 11A

Human Clinical Trial of the Effect of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxide(Compound 100A) on Necrotizing Enterocolitis (NEC)

Objective:

The aim of a 6 month study is to show evidence of the efficacy(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideon reduction of incidence and severity of NEC in very low birth weightinfants.

Study Design:

This is a 6 month study. A cohort of 1000 patients is divided intoplacebo group and drug treated groups. An optimized dose of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxidewill be administered with milk or formula.

Inclusion Criteria are: Infants with birth weight less than 1500 grams;gestational age 24 to 33 weeks; diagnosed with necrotizing enterocolitisbased on Bell stage II or greater; or bowel perforation as evidenced byfree intraperitoneal air seen on abdominal radiograph, stool, bile, orpus found at paracentesis or clinical evidence of perforation in thejoint opinion of the attending surgeon or neonatologist.

Exclusion Criteria are: Healthy infants, major congenital abnormalities,chronic disorders of other organs, evidence of gastrointestinal anomaly.

Patient Response to(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideTherapy

Patient response is assessed via primary and secondary outcome measures.

Primary Outcome Measures:

Disease mortality, disease progression, treatment failure, time torecovery.

Secondary Outcome Measures:

Inflammatory cytokine concentration, duration of inflammation, weightgain, length of hospital stay.

Blood Sampling

Serial blood is drawn by direct vein puncture before and afteradministration of Compound 100 analog. Venous blood samples (2 mL) fordetermination of serum concentrations are obtained at about 10 minutesprior to dosing and at approximately the following times after dosing:days 1, 2, 3, 4, 5, 6, 7, 14 and every 14 days later. Each serum sampleis divided into two aliquots. All serum samples are stored at −20° C.Serum samples are shipped on dry ice.

Pharmacokinetics:

Patients undergo plasma/serum sample collection for pharmacokineticevaluation before beginning treatment and at days 1, 2, 3, 4, 5, 6, 7,14 and every 14 days later. Pharmacokinetic parameters are calculated bymodel independent methods on a Digital Equipment Corporation VAX 8600computer system using the latest version of the BIOAVL software. Thefollowing pharmacokinetics parameters are determined: peak serumconcentration (C_(max)); time to peak serum concentration (t_(max));area under the concentration-time curve (AUC) from time zero to the lastblood sampling time (AUC₀₋₇₂) calculated with the use of the lineartrapezoidal rule; and terminal elimination half-life (t_(1/2)), computedfrom the elimination rate constant. The elimination rate constant isestimated by linear regression of consecutive data points in theterminal linear region of the log-linear concentration-time plot. Themean, standard deviation (SD), and coefficient of variation (CV) of thepharmacokinetic parameters are calculated for each treatment.

Statistical Analysis:

The association of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideand reduction of necrotizing enterocolitis is analyzed by χ2 test.

Example 11B

A Human Clinical Trial similar to the trial in Example 11A is conductedto determine the effect of1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate (Compound 100B) on reduction of necrotizingenterocolitis.

Example 11C

A Human Clinical Trial similar to the trial in Example 11A is conductedto determine the effect of1,1-Dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N—[(R)-α-[N-(2-sulphoethyl)carbamoyl]-4-hydroxybenzyl]carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepineon reduction of necrotizing enterocolitis.

Example 11D

A Human Clinical Trial similar to the trial in Example 11A is conductedto determine the effect of1,1-Dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N—[(R)-α-[N—((S)-1-carboxy-2-(R)-hydroxypropyl)carbamoyl]-4-hydroxybenzyl]carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepineon reduction of necrotizing enterocolitis.

Example 11E

A Human Clinical Trial similar to the trial in Example 11A is conductedto determine the effect of1,1-Dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N—[(R)-α-[N—((S)-1-carboxy-2-methylpropyl)carbamoyl]-4-hydroxybenzyl]carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepineon reduction of necrotizing enterocolitis.

Example 11F

A Human Clinical Trial similar to the trial in Example 11A is conductedto determine the effect of1-[[5-[[3-[(3S,4R,5R)-3-butyl-7-(dimethylamino)-3-ethyl-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5yl]phenyl]amino]-5-oxopentyl]amino]-1-deoxy-D-glucitolon reduction of necrotizing enterocolitis.

Example 11G

A Human Clinical Trial similar to the trial in Example 11A is conductedto determine the effect ofPotassium((2R,3R,4S,5R,6R)-4-benzyloxy-6-{3-[3-((3S,4R,5R)-3-butyl-7-dimethylamino-3-ethyl-4-hydroxy-1,1-dioxo-2,3,4,5-tetrahydro-1H-benzo[b]thiepin-5-yl)-phenyl]-ureido}-3,5-dihydroxy-tetrahydro-pyran-2-ylmethyl)sulphateethanolate, hydrate on reduction of necrotizing enterocolitis.

Example 11H

A Human Clinical Trial similar to the trial in Example 11A is conductedto determine the effect of SD-5613 (Pfizer, a.k.a. 97-G) on reduction ofnecrotizing enterocolitis.

Example 11I

A Human Clinical Trial similar to the trial in Example 11A is conductedto determine the effect of combination of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideand bile acids or mimetics on reduction of necrotizing enterocolitis.

Example 11J

A Human Clinical Trial similar to the trial in Example 11A is conductedto determine the effect of combination of1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate (Compound 100B) bile acids or mimetics on reduction ofnecrotizing enterocolitis.

Example 11K

A Human Clinical Trial similar to the trial in Example 11A is conductedto determine the effect of combination of1,1-Dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N—[(R)-α-[N-(2-sulphoethyl)carbamoyl]-4-hydroxybenzyl]carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepinebile acids or mimetics on reduction of necrotizing enterocolitis.

Example 11L

A Human Clinical Trial similar to the trial in Example 11A is conductedto determine the effect of combination of1,1-Dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N—[(R)-α-[N—((S)-1-carboxy-2-(R)-hydroxypropyl)carbamoyl]-4-hydroxybenzyl]carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepinebile acids or mimetics on reduction of necrotizing enterocolitis.

Example 11M

A Human Clinical Trial similar to the trial in Example 11A is conductedto determine the effect of combination of1,1-Dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N—[(R)-α-[N—((S)-1-carboxy-2-methylpropyl)carbamoyl]-4-hydroxybenzyl]carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepinebile acids or mimetics on reduction of necrotizing enterocolitis.

Example 11N

A Human Clinical Trial similar to the trial in Example 1 IA is conductedto determine the effect of combination of1-[[5-[[3-[(3S,4R,5R)-3-butyl-7-(dimethylamino)-3-ethyl-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5yl]phenyl]amino]-5-oxopentyl]amino]-1-deoxy-D-glucitolbile acids or mimetics on reduction of necrotizing enterocolitis.

Example 11O

A Human Clinical Trial similar to the trial in Example 11A is conductedto determine the effect of combination ofPotassium((2R,3R,4S,5R,6R)-4-benzyloxy-6-{3-[3-((3S,4R,5R)-3-butyl-7-dimethylamino-3-ethyl-4-hydroxy-1,1-dioxo-2,3,4,5-tetrahydro-1H-benzo[b]thiepin-5-yl)-phenyl]-ureido}-3,5-dihydroxy-tetrahydro-pyran-2-ylmethyl)sulphateethanolate, hydrate bile acids or mimetics on reduction of necrotizingenterocolitis.

Example 11P

A Human Clinical Trial similar to the trial in Example 11A is conductedto determine the effect of combination of SD-5613 (Pfizer, a.k.a. 97-G)bile acids or mimetics on reduction of necrotizing enterocolitis.

Example 12A

Human Clinical Trial of the effect of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxide(Compound 100A) on satiety and reduction in food intake, plasma glucoseand insulin levels

Objective:

The aim of the 8 week study is to show evidence of the efficacy(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideto induce satiety and to reduce food intake. A secondary objective ofthe study is to determine the effect of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideon food consumption (amount and composition) during a free meal(dinner); wellness after one week supplementation; the intermealinterval; and body weight and waist-hip ratio (WHR).

Study Design:

This is an 8 week study. A cohort of 500 patients is divided intoplacebo group and drug treated groups. 50 mg of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxidewill be administered 30 minutes prior to ingestion of lunch and prior toingestion of dinner for a total of 100 mg of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideper day.

Inclusion Criteria are: A BMI≥27, 70 years of age or less, waistcircumference ≥102 cm for men or ≥88 cm for women, eating disorders,diabetes

Exclusion Criteria are: Stroke/MI/unstable cardiovascular disease within6 months, clinically significant renal, hepatic or psychiatric disease,participation in a formal weight loss program or lifestyle intervention,glaucoma or intraocular pressure, use of any antidiabetic medication.

Blood Sampling

Serial blood is drawn by direct vein puncture before and afteradministration of Compound 100 analog. Venous blood samples (5 mL) fordetermination of serum concentrations are obtained at about 10 minutesprior to dosing and at approximately the following times after dosing:days 1, 2, 3, 4, 5, 6, 7, and 14. Each serum sample is divided into twoaliquots. All serum samples are stored at −20° C. Serum samples areshipped on dry ice.

Pharmacokinetics:

Patients undergo plasma/serum sample collection for pharmacokineticevaluation before beginning treatment and at days 1, 2, 3, 4, 5, 6, 7,and 14. Pharmacokinetic parameters are calculated by model independentmethods on a Digital Equipment Corporation VAX 8600 computer systemusing the latest version of the BIOAVL software. The followingpharmacokinetics parameters are determined: peak serum concentration(C_(max)); time to peak serum concentration (t_(max)); area under theconcentration-time curve (AUC) from time zero to the last blood samplingtime (AUC₀₋₇₂) calculated with the use of the linear trapezoidal rule;and terminal elimination half-life (t_(1/2)), computed from theelimination rate constant. The elimination rate constant is estimated bylinear regression of consecutive data points in the terminal linearregion of the log-linear concentration-time plot. The mean, standarddeviation (SD), and coefficient of variation (CV) of the pharmacokineticparameters are calculated for each treatment.

Patient Response to(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideTherapy

Patient response is assessed via weekly hospital visits, weight andwaist measurements and routine blood tests.

Primary Outcome Measures: Satiety scores (visual analog scores),wellness after one week supplementation.

Secondary Outcome Measures: Demonstrate an improvement over placebo inabsolute weight loss and waist to hip ratio. Demonstrate an improvementin quality of life.

Example 12B

A Human Clinical Trial similar to the trial in Example 12A is conductedto determine the effect of1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate (Compound 100B) on satiety and reduction in foodintake, plasma glucose and insulin level

Example 13

Human Phase I Clinical Trial of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxidealone or in combination with sitagliptin on satiety and reduction infood intake, plasma glucose and insulin levels

Aim of the Study:

A phase I dose-escalation study

Patient Population:

Obese type II diabetic

Study Design:

Randomized, double-blind, cross-over, single-dose placebo controlled

Ten subjects will be administered a single dose of sitagliptin tenminutes before administration of a single dose of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideprior to ingestion of food. A second group of ten subjects will beadministered a single dose of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxide30 minutes prior to ingestion of food. Rising dose levels of the ASBTI(1, 3, 10 and 30 mg/kg) will be administered at each visit. Each visitto test facility will be at least 72 hours apart.

A buffet style breakfast will be presented and ad-libitum food intakewill be measured. Patients will record meal-related satiation, orhunger, and nausea on a chart. Satiogenic gut peptides will be measuredpost-prandially from plasma analytes. Insulin, glucose, GLP-1 and PYYlevels in blood/plasma will be monitored for 24 hours. Subjects will beasked to record food intake for 72 hours after each dose.

Endpoints:

Safety and tolerability, pharmacodynamics, food intake 24 hours afterdosing.

Example 14

A Human Phase II Clinical Trial of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxide(Compound 100A) alone or in combination with sitagliptin and effect onobesity or diabetes

Phase 2A:

This will be a 16 week double-blind, parallel-group study. A single doseof sitagliptin in combination with(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxidewill be administered as described above. A second group of subjects willbe administered a single dose of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxide30 minutes prior to ingestion of food. A buffet style breakfast will bepresented as described above and satiety and food intake will bemeasured.

Phase 2A:

This will be a 6-week, single-blind, parallel-group study carried out asdescribed above. Glucose levels in blood/plasma, body composition and acomprehensive assessment of food intake will be carried for the durationof the study.

Phase 2B:

This will be a 16-week, double-blind, parallel group study dose-rangingstudy carried out as described above. Safety, dose-tolerance, glucoselevels in blood/plasma, weight loss and HBA1c will be monitored for theduration of the study.

Example 15

A Human Phase II Clinical Trial similar to the trial in Example 14 isconducted to determine the effect of1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate (Compound 100B) alone or in combination withsitagliptin and effect on obesity or diabetes

Example 16

A Human Phase II Clinical Trial similar to the trial in Example 14 isconducted to determine the effect of compound 4 alone or in combinationwith sitagliptin and effect on obesity or diabetes

Example 17

Human Phase I Clinical Trial of1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate (Compound 100B) in combination with sitagliptin onsatiety and reduction in food intake, plasma glucose and insulin level

Aim of the Study:

A phase I dose-escalation study

Patient Population:

Obese type II diabetic

Study Design:

Randomized, double-blind, cross-over, single-dose placebo controlled

Ten subjects will be administered a single dose of sitagliptin tenminutes before administration of a single dose of1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate prior to ingestion of food. A second group of tensubjects will be administered a single dose of1-[4-[4-[(4R,5R)-3,3-dibutyl-7-(dimethylamino)-2,3,4,5-tetrahydro-4-hydroxy-1,1-dioxido-1-benzothiepin-5-yl]phenoxy]butyl]4-aza-1-azoniabicyclo[2.2.2]octanemethane sulfonate 30 minutes prior to ingestion of food. Rising doselevels of the ASBTI (1, 3, 10 and 30 mg/kg) will be administered at eachvisit. Each visit to test facility will be at least 72 hours apart.

A buffet style breakfast will be presented and ad-libitum food intakewill be measured. Patients will record meal-related satiation, orhunger, and nausea on a chart. Satiogenic gut peptides will be measuredpost-prandially from plasma analytes. Insulin, glucose, GLP-1 and PYYlevels in blood/plasma will be monitored for 24 hours. Subjects will beasked to record food intake for 72 hours after each dose.

Endpoints:

Safety and tolerability, pharmacodynamics, food intake 24 hours afterdosing.

Example 18

A Human Phase I Clinical Trial similar to the trial in Example 17 isconducted to determine the effect of compound 2 in combination withsitagliptin on satiety and reduction in food intake, plasma glucose andinsulin level

Example 19

Human Clinical Trial of the Effect of Compound 4 on Satiety andReduction in Food Intake

Objective:

The aim of the 8 week study is to show evidence of the efficacy ofcompound 4 to induce satiety and to reduce food intake. A secondaryobjective of the study is to determine the effect of compound 4 on foodconsumption (amount and composition) during a free meal (dinner);wellness after one week supplementation; the intermeal interval; andbody weight and waist-hip ratio (WHR).

Study Design:

This is an 8 week study. A cohort of 500 patients is divided intoplacebo group and drug treated groups. 50 mg of compound 4 will beadministered 30 minutes prior to ingestion of lunch and prior toingestion of dinner for a total of 100 mg of compound 4 per day.

Inclusion Criteria are: A BMI≥27, 70 years of age or less, waistcircumference ≥102 cm for men or ≥88 cm for women.

Exclusion Criteria are: Stroke/MI/unstable cardiovascular disease within6 months, clinically significant renal, hepatic or psychiatric disease,participation in a formal weight loss program or lifestyle intervention,glaucoma or intraocular pressure, use of any antidiabetic medication,previous bariatric surgery

Blood Sampling

Serial blood is drawn by direct vein puncture before and afteradministration of Compound 100 analog. Venous blood samples (5 mL) fordetermination of serum concentrations are obtained at about 10 minutesprior to dosing and at approximately the following times after dosing:days 1, 2, 3, 4, 5, 6, 7, and 14. Each serum sample is divided into twoaliquots. All serum samples are stored at −20° C. Serum samples areshipped on dry ice.

Pharmacokinetics:

Patients undergo plasma/serum sample collection for pharmacokineticevaluation before beginning treatment and at days 1, 2, 3, 4, 5, 6, 7,and 14. Pharmacokinetic parameters are calculated by model independentmethods on a Digital Equipment Corporation VAX 8600 computer systemusing the latest version of the BIOAVL software. The followingpharmacokinetics parameters are determined: peak serum concentration(C_(max)); time to peak serum concentration (t_(max)); area under theconcentration-time curve (AUC) from time zero to the last blood samplingtime (AUC₀₋₇₂) calculated with the use of the linear trapezoidal rule;and terminal elimination half-life (t_(1/2)), computed from theelimination rate constant. The elimination rate constant is estimated bylinear regression of consecutive data points in the terminal linearregion of the log-linear concentration-time plot. The mean, standarddeviation (SD), and coefficient of variation (CV) of the pharmacokineticparameters are calculated for each treatment.

Patient Response to Compound 4 Therapy:

Patient response is assessed via weekly hospital visits, weight andwaist measurements and routine blood tests.

Primary Outcome Measures:

Satiety scores (visual analog scores), wellness after one weeksupplementation.

Secondary Outcome Measures:

Demonstrate an improvement over placebo in absolute weight loss andwaist to hip ratio. Demonstrate an improvement in quality of life.

Example 20

Human Clinical Trial of the effect of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxide(Compound 100A) on risk of pancreatic cancer

Objective:

The aim of a 1 year study is to show evidence of the efficacy(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideon reduction of risk of pancreatic cancer.

Study Design:

This is a 1 year study. A cohort of 1000 patients is divided intoplacebo group and drug treated groups. 50 mg of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxidewill be administered 30 minutes prior to ingestion of lunch and prior toingestion of dinner for a total of 100 mg of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideper day.

Inclusion Criteria are: 70 years of age or less, waist circumference≥102 cm for men or ≥88 cm for women, pancreatic ductal adenocarcinoma.

Exclusion Criteria are: Stroke/MI/unstable cardiovascular disease within6 months, clinically significant renal, hepatic or psychiatric disease,participation in a formal weight loss program or lifestyle intervention,glaucoma or intraocular pressure, use of any antidiabetic medication,prior history with cancer.

Patient Response to(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideTherapy

Patient response is assessed via weekly hospital visits, weight andwaist measurements and routine blood tests.

Blood Sampling

Serial blood is drawn by direct vein puncture before and afteradministration of Compound 100 analog. Venous blood samples (5 mL) fordetermination of serum concentrations are obtained at about 10 minutesprior to dosing and at approximately the following times after dosing:days 1, 2, 3, 4, 5, 6, 7, 14 and every 14 days later. Each serum sampleis divided into two aliquots. All serum samples are stored at −20° C.Serum samples are shipped on dry ice.

Pharmacokinetics:

Patients undergo plasma/serum sample collection for pharmacokineticevaluation before beginning treatment and at days 1, 2, 3, 4, 5, 6, 7,14 and every 14 days later. Pharmacokinetic parameters are calculated bymodel independent methods on a Digital Equipment Corporation VAX 8600computer system using the latest version of the BIOAVL software. Thefollowing pharmacokinetics parameters are determined: peak serumconcentration (C_(max)); time to peak serum concentration (t_(max));area under the concentration-time curve (AUC) from time zero to the lastblood sampling time (AUC₀₋₇₂) calculated with the use of the lineartrapezoidal rule; and terminal elimination half-life (t_(1/2)), computedfrom the elimination rate constant. The elimination rate constant isestimated by linear regression of consecutive data points in theterminal linear region of the log-linear concentration-time plot. Themean, standard deviation (SD), and coefficient of variation (CV) of thepharmacokinetic parameters are calculated for each treatment.

Patient Response to Compound 4 Therapy:

Patient response is assessed via weekly hospital visits, weight andwaist measurements and routine blood tests.

Statistical Analysis:

The association of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideand risk of pancreatic cancer is analyzed in multivariable logisticregression models including age, sex, race, smoking, alcohol, BMI,family history of cancer, duration of diabetes, and insulin use. Tocontrol for reversal causality due to pancreatic cancer-caused diabetes,risk of pancreatic cancer is estimated after exclusion of those withduration of diabetes ≤2 years. HbA1c level (≤7% or ≥7%), a marker ofglycemic control, is compared between placebo and users of(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideby χ2 test. The demographic and risk factors as well as duration ofdiabetes and insulin use are compared between(−)-(3R,5R)-trans-3-butyl-3-ethyl-2,3,4,5-tetrahydro-7,8-dimethoxy-5-phenyl-1,4-benzothiazepine1,1-dioxideusers and placebo by χ2 test.

Example 21

Rectal Foams

a) 500 mM Sodium Taurocholate

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer and turboemulsifier 26.88 grams of sodium taurocholate, 0.25grams of potassium metabisulphite, 0.3 grams EDTA (disodium salt), 0.38grams of sodium benzoate and 0.2 grams of xanthan gum are dissolved in100 mL of purified water. While stirring, 4 grams of Polysorbate 20 and4 grams of Polyglycol 300 isostearate are added and stirring iscontinued for 15 minutes. The suspension is then pumped into an aerosolcans and is immediately sealed by clinching the dispenser valve. The canis then pressurized by pumping 6.5 grams of Freon 12 and 3.5 grams ofFreon 114 into the can.

b) 500 mM Sodium Glycocholate

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer and turboemulsifier 24.38 grams of sodium glycocholate, 0.25grams of potassium metabisulphite, 0.3 grams EDTA (disodium salt), 0.38grams of sodium benzoate and 0.2 grams of xanthan gum are dissolved in100 mL of purified water. While stirring, 4 grams of Polysorbate 20 and4 grams of Polyglycol 300 isostearate are added and stirring iscontinued for 15 minutes. The suspension is then pumped into an aerosolcans and is immediately sealed by clinching the dispenser valve. The canis then pressurized by pumping 6.5 grams of Freon 12 and 3.5 grams ofFreon 114 into the can.

c) No Bile Salt (Control)

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer and turboemulsifier 0.25 grams of potassium metabisulphite, 0.3grams EDTA (disodium salt), 0.38 grams of sodium benzoate and 0.2 gramsof xanthan gum are dissolved in 100 mL of purified water. Whilestirring, 4 grams of Polysorbate 20 and 4 grams of Polyglycol 300isostearate are added and stirring is continued for 15 minutes. Thesuspension is then pumped into an aerosol cans and is immediately sealedby clinching the dispenser valve. The can is then pressurized by pumping6.5 grams of Freon 12 and 3.5 grams of Freon 114 into the can.

Analysis of Food Intake

The sodium taurocholate rectal foam described above is rectallyadministered to 5 conscious overnight-fasted subjects (e.g., SpragueDawley rats) and the control rectal foam without the sodium taurocholateis rectally administered to 5 conscious overnight-fasted subjects (e.g.,Sprague Dawley rats). Each subject is exposed to pre-weighed food andthe cumulative consumption of the food is determined over a 6 hourperiod by weighing the food every 30 minutes.

The sodium glycocholate rectal foam described above is rectallyadministered to 5 conscious overnight-fasted subjects (e.g., SpragueDawley rats) and the control rectal foam without the sodium taurocholateis rectally administered to 5 conscious overnight-fasted subject (e.g.,Sprague Dawley rats). Each subject is exposed to pre-weighed food andthe cumulative consumption of the food is determined over a 6 hourperiod by weighing the food every 30 minutes.

Example 22

Rectal Enemas

a) 500 mM Sodium Taurocholate

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer 26.88 grams of sodium taurocholate, 0.25 grams of potassiummetabisulphite, 0.3 grams EDTA (disodium salt), 0.38 grams of sodiumbenzoate are dissolved in 100 mL of purified water and stirring iscontinued for 10 minutes. The solution is then pulled into a syringe.

b) 500 mM Sodium Glycocholate

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer and turboemulsifier 24.38 grams of sodium glycocholate, 0.25grams of potassium metabisulphite, 0.3 grams EDTA (disodium salt), 0.38grams of sodium benzoate are dissolved in 100 mL of purified water andstirring is continued for 10 minutes. The solution is then pulled into asyringe.

c) No Bile Salt (Control)

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer and turboemulsifier 0.25 grams of potassium metabisulphite, 0.3grams EDTA (disodium salt), 0.38 grams of sodium benzoate are dissolvedin 100 mL of purified water and stirring is continued for 10 minutes.The solution is then pulled into a syringe.

Analysis of Food Intake

The sodium taurocholate rectal enema described above is rectallyadministered to 5 conscious overnight-fasted subjects (e.g., SpragueDawley rats) and the control rectal enema without the sodiumtaurocholate is rectally administered to 5 conscious overnight-fastedsubjects (e.g., Sprague Dawley rats). Each subject is exposed topre-weighed food and the cumulative consumption of the food isdetermined over a 6 hour period by weighing the food every 30 minutes.

The sodium glycocholate rectal enema described above is rectallyadministered to 5 conscious overnight-fasted subjects (e.g., SpragueDawley rats) and the control rectal enema without the sodiumtaurocholate is rectally administered to 5 conscious overnight-fastedsubjects (e.g., Sprague Dawley rats). Each subject is exposed topre-weighed food and the cumulative consumption of the food isdetermined over a 6 hour period by weighing the food every 30 minutes.

Example 23

Rectal Suppositories

a) Sodium Taurocholate

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer 2.69 grams of sodium taurocholate and 0.1 grams of methylcellulose are added to 10 grams of higher saturated fatty acidtriglycerides (Witepsol™S55; Dynamic Novel Aktiengesellschaft, WestGermany) and the combination is melted at 50 C and stirred. While thecomposition is a liquid it is filled into suppository containers forrats (50 mg per container) and then quenched in ice-water.

b) 500 mM Sodium Glycocholate

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer 2.69 grams of sodium glycocholate and 0.1 grams of methylcellulose are added to 10 grams of higher saturated fatty acidtriglycerides (Witepsol™S55; Dynamic Novel Aktiengesellschaft, WestGermany) and the combination is melted at 50 C and stirred. While thecomposition is a liquid it is filled into suppository containers forrats (50 mg per container) and then quenched in ice-water.

c) No Bile Salt (Control)

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer 0.1 grams of methyl cellulose is added to 10 grams of highersaturated fatty acid triglycerides (Witepsol™S55; Dynamic NovelAktiengesellschaft, West Germany) and the combination is melted at 50 Cand stirred. While the composition is a liquid it is filled intosuppository containers for rats (50 mg per container) and then quenchedin ice-water.

Analysis of Food Intake

The sodium taurocholate rectal suppository described above is rectallyadministered to 5 conscious overnight-fasted subjects (e.g., SpragueDawley rats) and the control rectal suppository without the sodiumtaurocholate is rectally administered to 5 conscious overnight-fastedsubjects (e.g., Sprague Dawley rats). Each subject is exposed topre-weighed food and the cumulative consumption of the food isdetermined over a 6 hour period by weighing the food every 30 minutes.

The sodium glycocholate rectal suppository described above is rectallyadministered to 5 conscious overnight-fasted subjects (e.g., SpragueDawley rats) and the control rectal suppository without the sodiumglycocholate is rectally administered to 5 conscious overnight-fastedsubjects (e.g., Sprague Dawley rats). Each subject is exposed topre-weighed food and the cumulative consumption of the food isdetermined over a 6 hour period by weighing the food every 30 minutes.

Example 24

Rectal Gels—Sodium Taurocholate/Control

a) 500 mM Sodium Taurocholate

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer 26.88 grams of sodium taurocholate and 1 gram of methylcellulose are dissolved in 100 mL of purified water and stirred for 15minutes. 6 syringes connected to gavage tubes were then each filled with3 mL of the composition.

b) No Bile Salt (Control)

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer 1 gram of methyl cellulose is dissolved in 100 mL of purifiedwater and stirred for 15 minutes. 5 syringes connected to gavage tubesare then each filled with 3 mL of the composition.

Analysis of Food Intake

The sodium taurocholate rectal gel described above was rectallyadministered to 6 conscious overnight-fasted subjects (e.g., SpragueDawley rats) and the control rectal gel without the sodium taurocholatewas rectally administered to 5 conscious overnight-fasted subjects(e.g., Sprague Dawley rats). Each rat was then exposed to pre-weighedfood and the cumulative consumption of the food was determined over a 6hour period by weighing the food after 30 minutes, 60 minutes, 120minutes, 240 minutes and 360 minutes.

Results

The cumulative food intake in rats instilled with taurocholatecontaining gels is reduced compared to rats instilled with gel alone(FIG. 5). FIG. 5 illustrates the individual food intake in rats withbile sat containing gel per rectum (dotted lines) or control vehicle(solid lines). The effect of the taurocholate on food intake is found tobe statistically significant.

Example 5

Rectal Gels—Sodium Taurcholate Dose Response

a) 50 mM Sodium Taurocholate

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer 2.688 grams of sodium taurocholate and 1 gram of methylcellulose are dissolved in 100 mL of purified water and stirred for 15minutes. 12 syringes connected to gavage tubes are then each filled with3 mL of the composition.

b) 150 mM Sodium Taurocholate

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer 8.066 grams of sodium taurocholate and 1 gram of methylcellulose are dissolved in 100 mL of purified water and stirred for 15minutes. 12 syringes connected to gavage tubes are then each filled with3 mL of the composition.

c) 500 mM Sodium Taurocholate

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer 26.88 grams of sodium taurocholate and 1 gram of methylcellulose are dissolved in 100 mL of purified water and stirred for 15minutes. 12 syringes connected to gavage tubes are then each filled with3 mL of the composition.

d) No Bile Salt (Control)

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer 1 gram of methyl cellulose is dissolved in 100 mL of purifiedwater and stirred for 15 minutes. 12 syringes connected to gavage tubesare then each filled with 3 mL of the composition.

Analysis of Food Intake

A 4×4 Latin Square design is used to monitor the effect of the differentconcentration of taurocholate on the food uptake of overnight-fastedSprague Dawley rats. Each concentration is tested in triplicate withfour rats used per replicate. Therefore, twelve rats were used for eachrectal gel composition (50 mM gel, 150 mM gel, 500 mM gel and thecontrol gel), and all rats received each of the 4 treatments. Followingrectal administration of the gels, each rat is then exposed topre-weighed food and the cumulative consumption of the food isdetermined over a 24 hour period by weighing the food after 30 minutes,1 hour, 2 hours, 4 hours, 6 hours, 8 hours and 24 hours.

Results

The cumulative food intake in rats instilled with taurocholatecontaining gels is observed to be dose dependently related to thequantity of taurocholate administered (FIG. 6). Moreover, thedifferences in food uptake are observed to be maintained for at least 24hours.

The ED50 for inhibition of food intake is obtained by plotting thenormalizing the dose response for the anorectic effect of taurocholateto the control data as a log of taurocholate dose vs the percentage ofcontrol cumulative intake (FIG. 7). The ED50 for inhibition of foodintake at 1 hour is 528 mM for 3 mL doses (1.6 mmol/rat) while for 24hour intake the ED50 is 7.2 mmol/rat. Surprisingly, the dose dependencyfor inhibition of intake is not different between 8 and 24 hours,thereby indicating the longevity of the anorectic response.

Example 26

Rectal Gels—Sodium Glycocholate/Control

a) 500 mM Sodium Glycocholate

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer 24.38 grams of sodium glucocholate and 1 gram of methylcellulose are dissolved in 100 mL of purified water and then stirred for15 minutes. 6 syringes connected to gavage tubes are then each filledwith 3 mL of the composition.

b) No Bile Salt (Control)

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer 1 gram of methyl cellulose is dissolved in 100 mL of purifiedwater and stirred for 15 minutes. 5 syringes connected to gavage tubesare then each filled with 3 mL of the composition.

Analysis of Food Intake

The sodium glycocholate rectal gel described above is rectallyadministered to 6 conscious overnight-fasted subjects (e.g., SpragueDawley rats) and the control rectal gel without the sodium glycocholateis rectally administered to 5 conscious overnight-fasted subjects (e.g.,Sprague Dawley rats). Each subject is then exposed to pre-weighed foodand the cumulative consumption of the food is determined over a 6 hourperiod by weighing the food after 30 minutes, 60 minutes, 120 minutes,240 minutes and 360 minutes.

Example 27

Rectal Gels—Sodium Glycocholate Dose Response

a) 50 mM Sodium Glycocholate

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer 2.44 grams of sodium glycocholate and 1 gram of methyl celluloseare dissolved in 100 mL of purified water and then stirred for 15minutes. 12 syringes connected to gavage tubes are then each filled with3 mL of the composition.

b) 150 mM Sodium Glycocholate

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer 7.32 grams of sodium glycocholate and 1 gram of methyl celluloseare dissolved in 100 mL of purified water and then stirred for 15minutes. 12 syringes connected to gavage tubes are then each filled with3 mL of the composition.

c) 500 mM Sodium Glycocholate

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer 24.38 grams of sodium glycocholate and 1 gram of methylcellulose are dissolved in 100 mL of purified water and then stirred for15 minutes. 12 syringes connected to gavage tubes are then each filledwith 3 mL of the composition.

d) No Bile Salt (Control)

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer 1 gram of methyl cellulose is dissolved in 100 mL of purifiedwater and then stirred for 15 minutes. 12 syringes connected to gavagetubes are then each filled with 3 mL of the composition.

Analysis of Food Intake

A 4×4 Latin Square design as used to monitor the effect of the differentconcentration of glycocholate on the food uptake of overnight-fastedSprague Dawley rats. Each concentration is tested in triplicate withfour rats used per replicate. Therefore, twelve rats are used for eachrectal gel composition (50 mM gel, 150 mM gel, 500 mM gel and thecontrol gel), with all rats receiving each of the 4 treatments.Following rectal administration of the gels, each rat is then exposed topre-weighed food and the cumulative consumption of the food isdetermined over a 24 hour period by weighing the food after 30 minutes,1 hour, 2 hours, 4 hours, 6 hours, 8 hours and 24 hours.

Example 28

Enteric Coated Tablets

a) 5 mg Sodium Taurocholate

Preparation Method:

Preparation of core: 5 mg sodium taurocholate, 25 mg microcrystallinecellulose, 20 mg mannitol, and 10 mg croscarmellose sodium are mixed ina Hobart Mixer for 15 minutes. The mixture is granulated with 20%polyvinyl pyrrolidone (4 mg) solution until optimum granulation isobtained. The granulation is dried overnight at 50° C. The granulationis then passed through a #30 mesh. The granulation is then blended with1 mg magnesium stearate. Using an F-Press ¼″ standard concave roundpunch, the granulation is compressed into a tablet. Preparation oferodible polymer layer and dual matrix tablets: 415 mg hydroxypropylmethylcellulose, 75 mg microcrystalline cellulose, and 6 mgpolyvinylpyrrolidone are uniformly mixed with a mortar. The powder mixis granulated with 50% v/v alcohol solution until optimum granulation isobtained. The granulation is dried overnight at 50° C. The granulationis then passed through a #40 mesh screen. The granulation is thenblended with 2.5 mg magnesium stearate. Using a Carver Press and a 7/16″standard concave round punch, half of the granulation is placed in thedie cavity, the core is then placed in the cavity and the other half ofthe granulation is placed in the die cavity. The mass is compressed to5,000 lbs to form the dual matrix tablet. Enteric coating: Using apropellar mixer, 42 g of hydroxypropyl methylcellulose phthalate and 4.2g of distilled acetylated monoglycerides are dissolved in 514 mL of amixture of a cetone and absolute alcohol (1:1). Using a spray system,the dual matrix tablets are then coated with the enteric coatingsolution. Approximately 60 mg of the coating material (dry basis) isapplied per tablet.

b) 500 mM Sodium Glycocholate

Preparation Method:

Preparation of core: 5 mg sodium glycocholate, 25 mg microcrystallinecellulose, 20 mg mannitol, and 10 mg croscarmellose sodium are mixed ina Hobart Mixer for 15 minutes. The mixture is granulated with 20%polyvinyl pyrrolidone (4 mg) solution until optimum granulation isobtained. The granulation is dried overnight at 50° C. The granulationis then passed through a #30 mesh. The granulation is then blended with1 mg magnesium stearate. Using an F-Press ¼″ standard concave roundpunch, the granulation is compressed into a tablet. Preparation oferodible polymer layer and dual matrix tablets: 415 mg hydroxypropylmethylcellulose, 75 mg microcrystalline cellulose, and 6 mgpolyvinylpyrrolidone are uniformly mixed with a mortar. The powder mixis granulated with 50% v/v alcohol solution until optimum granulation isobtained. The granulation is dried overnight at 50° C. The granulationis then passed through a #40 mesh screen. The granulation is thenblended with 2.5 mg magnesium stearate. Using a Carver Press and a 7/16″standard concave round punch, half of the granulation is placed in thedie cavity, the core is then placed in the cavity and the other half ofthe granulation is placed in the die cavity. The mass is compressed to5,000 lbs to form the dual matrix tablet. Enteric coating: Using apropellar mixer, 42 g of hydroxypropyl methylcellulose phthalate and 4.2g of distilled acetylated monoglycerides are dissolved in 514 mL of amixture of a cetone and absolute alcohol (1:1). Using a spray system,the dual matrix tablets are then coated with the enteric coatingsolution. Approximately 60 mg of the coating material (dry basis) isapplied per tablet.

c) No Bile Salt (Control)

Preparation Method:

Preparation of core: 25 mg microcrystalline cellulose, 20 mg mannitol,and 10 mg croscarmellose sodium are mixed in a Hobart Mixer for 15minutes. The mixture is granulated with 20% polyvinyl pyrrolidone (4 mg)solution until optimum granulation is obtained. The granulation is driedovernight at 50° C. The granulation is then passed through a #30 mesh.The granulation is then blended with 1 mg magnesium stearate. Using anF-Press ¼″ standard concave round punch, the granulation is compressedinto a tablet. Preparation of erodible polymer layer and dual matrixtablets: 415 mg hydroxypropyl methylcellulose, 75 mg microcrystallinecellulose, and 6 mg polyvinylpyrrolidone are uniformly mixed with amortar. The powder mix is granulated with 50% v/v alcohol solution untiloptimum granulation is obtained. The granulation is dried overnight at50° C. The granulation is then passed through a #40 mesh screen. Thegranulation is then blended with 2.5 mg magnesium stearate. Using aCarver Press and a 7/16″ standard concave round punch, half of thegranulation is placed in the die cavity, the core is then placed in thecavity and the other half of the granulation is placed in the diecavity. The mass is compressed to 5,000 lbs to form the dual matrixtablet. Enteric coating: Using a propellar mixer, 42 g of hydroxypropylmethylcellulose phthalate and 4.2 g of distilled acetylatedmonoglycerides are dissolved in 514 mL of a mixture of a cetone andabsolute alcohol (1:1). Using a spray system, the dual matrix tabletsare then coated with the enteric coating solution. Approximately 60 mgof the coating material (dry basis) is applied per tablet.

Analysis of Food Intake

The sodium taurocholate tablet described above (or, when necessary giventhe size and identity of a subject, a similar tablet having anappropriate dose and size) is orally administered to 5 consciousovernight-fasted subjects and the control tablet without the sodiumtaurocholate is orally administered to 5 conscious overnight-fastedsubjects. Each subject is exposed to pre-weighed food and the cumulativeconsumption of the food is determined over a 6 hour period by weighingthe food every 30 minutes.

The sodium glycocholate tablet described above (or, when necessary giventhe size and identity of a subject, a similar tablet having anappropriate dose and size) is orally administered to 5 consciousovernight-fasted subjects and the control tablet without the sodiumtaurocholate is orally administered to 5 conscious overnight-fastedsubjects. Each subject is exposed to pre-weighed food and the cumulativeconsumption of the food is determined over a 6 hour period by weighingthe food every 30 minutes.

Example 29

Absorption Inhibitors

a) Control. 500 mM Sodium Taurocholate

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer and turboemulsifier 26.88 grams of sodium taurocholate, 0.25grams of potassium metabisulphite, 0.3 grams EDTA (disodium salt) and0.38 grams of sodium benzoate dissolved in 100 mL of purified water.While stirring, 4 grams of Polysorbate 20 and 4 grams of Polyglycol 300isostearate are added and stirring is continued for 15 minutes. Thesuspension is then pumped into an aerosol cans and is immediately sealedby clinching the dispenser valve. The can is then pressurized by pumping6.5 grams of Freon 12 and 3.5 grams of Freon 114 into the can.

b) 500 mM Sodium Taurocholate+Candidate Absorption Inhibitor

Preparation Method:

Using a stainless steel dissolving vessel fitted with a propellerstirrer and turboemulsifier 26.88 grams of sodium taurocholate, 0.25grams of potassium metabisulphite, 0.3 grams EDTA (disodium salt), 0.38grams of sodium benzoate and between 0.01 grams and 20 grams of acandidate absorption inhibitor are dissolved in 100 mL of purifiedwater. While stirring, 4 grams of Polysorbate 20 and 4 grams ofPolyglycol 300 isostearate are added and stirring is continued for 15minutes. The suspension is then pumped into an aerosol cans and isimmediately sealed by clinching the dispenser valve. The can is thenpressurized by pumping 6.5 grams of Freon 12 and 3.5 grams of Freon 114into the can.

Analysis of Absorption Inhibition

The foams described above are rectally administered to 5 consciousovernight-fasted subjects (e.g., Sprague Dawley rats). Each subject isexposed to pre-weighed food and the cumulative consumption of the foodis determined over a 6 hour period by weighing the food every 30minutes. Food intake is compared between the groups. The candidateabsorption inhibitor inhibits absorption of the enteroendocrine peptidesecretion enhancing agent (in this case sodium taurocholate) when theenteroendocrine peptide secretion enhancing agent is able to interactwith the L-cells for a longer period of time (i.e., when it is notsystemically absorbed), thereby reducing food intake when compared tothe control formulation.

Alternatively, the ability of the absorption inhibitor to inhibit theabsorption of the enteroendocrine peptide secretion enhancing agent (inthis case sodium taurocholate) across the colon and/or rectum mucosa isdetermined by measuring the systemic concentration of enteroendocrinepeptide secretion enhancing agent. Systemic concentration ofenteroendocrine peptide secretion enhancing agent is measured prior toadministration and at a time following administration of theenteroendocrine peptide secretion enhancing agent (e.g., after onehour). Decreased systemic concentration of the enteroendocrine peptidesecretion enhancing agent indicate that the candidate absorptioninhibitor inhibits the absorption of the enteroendocrine peptidesecretion enhancing agent.

Example 30

Method of Inhibiting Food Intake

Studies are undertaken after an overnight fast following administration(8-10 hours prior to bile salt) of 100 mg Januvia (sitagliptin), a DDP-4inhibitor that protects GLP-1 from inactivation by proteolysis. Doseescalation involves half-log increases in dose as tolerated. Planneddoses are 1, 2, 7 and 20 mmoles of deoxycholic acid. Each dose isadministered on a different day with at least three days between doses.

Subjects undergo complete physical examination with laboratoryinvestigations including complete blood count, glucose, fasting lipids,liver function tests, urea and electrolytes, prothrombin time andpartial thromboplastin time, haemoglobin A_(1C), and urinalysis.Subjects are excluded if the fasting blood glucose is >300 mg/dl, if thehaemoglobin A_(1C) is >11%, or if there are abnormal liver functiontests (such as transaminase levels >3× the upper limit of normal.

Subjects on oral medication will not receive such medication during thefasting period prior to the study or during the study period.

Subjects are studied on four separate occasions, e.g., at weeklyintervals with at least three days gap between the different occasions.Subjects are studied after an overnight (10 hour) fast. Ten hours beforethe procedure, each subject will take 100 mg oral Januvia (sitagliptin),a DDP-4 inhibitor to protect GLP-1 (7-36) amide from degradation in thecirculation. An indwelling catheter (Intracath) is placed in a forearmvein for blood sampling. An initial basal blood sample (5 ml) is takenand the catheter kept patent with normal saline. Blood samples are takeninto EDTA Vacutainers (purple top). After 15 minutes a second bloodsample (5 ml) will be collected. Placebo (vehicle) or sodiumdeoxycholate at doses of 1, 2, 7 and 20 mmoles, incorporated into 20 mLor 60 mL of 1% carboxymethyl cellulose is placed per rectum by syringe(type) over a period of one minute. Further 5 ml blood samples arecollected 10, 20, 30, 40, 50 and 60 minutes after instillation of theplacebo or deoxycholate.

In plasma samples from each time point we will measure glucose, insulin,GLP-1 (active) and PYY (total), using assay kits (Linco) from theMillipore Corporation. Measures of these hormones provide an endpointfor determining efficacy in producing reduced food intake.

In addition, measured is food intake for a two hour period after the endof the infusion study. During this period, the subjects are offered abuffet lunch with food in such excess that all appetites will besatisfied. The amount of food is quantified preprandially andpostprandially and the caloric intake calculated. Before, at the end ofthe infusion study, and two hours after presentation of a buffet lunch,appetite ratings are made on a 100 mm visual analogue scale (highervalues indicating greater appetite) with the text expressing the mostpositive and most negative ratings at each end of the scale.

Example 31

In certain instances, placing bile salts or other enteroendocrinepeptide enhancing agents into the rectum has several advantages andprovides substantial information on the whole process of releasing thedistal gut hormones, GLP-1, oxyntomodulin and PYY. In our human studieswe have demonstrated the following:

-   -   Dose-responsive increase in GLP-1 and PYY levels in the        bloodstream.    -   Consequent increase in insulin secretion and reduction in        glucose levels.    -   Dose-responsive and substantial reduction in food intake.    -   Elevation of high local concentrations of bile salt in the        rectum without diarrhea.

Example 32

Studies were undertaken after an overnight fast following administration(8-10 hours prior to bile salt) of 100 mg sitagliptin, a DPP-4 inhibitorthat protects GLP-1 from inactivation by proteolysis. Dose escalationinvolved half-log increases in dose of taurocholic acid. The doses usedwere 0.66, 2.0, 6.66 and 20.0 mmoles (358, 1075, 3584, 10,754 mg) oftaurocholic acid. The taurocholic acid was administered into the rectumby syringe in a total volume of 20 ml of 1% carboxymethyl cellulose gel.Each dose was administered on a different day with at least three daysbetween doses. FIG. 9 illustrates the increase of circulating GLP-1levels following rectal administration of taurocholic acid. FIG. 10illustrates the increase of circulating PYY levels following rectaladministration of taurocholic acid

Example 33

Objective:

The aim of a 1 year study is to show evidence of the efficacy SodiumTaurocholate on reduction of risk of pancreatic cancer.

Study Design:

This is a 1 year study. A cohort of 1000 patients is divided intoplacebo group and drug treated groups. 50 mg of Sodium taurocholate willbe rectally administered 30 minutes prior to ingestion of lunch andprior to ingestion of dinner for a total of 100 mg of Sodiumtaurocholate per day.

Inclusion Criteria are: 70 years of age or less, waist circumference≥102 cm for men or ≥88 cm for women, pancreatic ductal adenocarcinoma.

Exclusion Criteria are: Stroke/MI/unstable cardiovascular disease within6 months, clinically significant renal, hepatic or psychiatric disease,participation in a formal weight loss program or lifestyle intervention,glaucoma or intraocular pressure, use of any antidiabetic medication,prior history with cancer.

Patient Response to Sodium Taurocholate Therapy:

Patient response is assessed via weekly hospital visits, weight andwaist measurements and routine blood tests.

Blood Sampling

Serial blood is drawn by direct vein puncture before and afteradministration of Compound 100 analog. Venous blood samples (5 mL) fordetermination of serum concentrations are obtained at about 10 minutesprior to dosing and at approximately the following times after dosing:days 1, 2, 3, 4, 5, 6, 7, 14 and every 14 days later. Each serum sampleis divided into two aliquots. All serum samples are stored at −20° C.Serum samples are shipped on dry ice.

Pharmacokinetics:

Patients undergo plasma/serum sample collection for pharmacokineticevaluation before beginning treatment and at days 1, 2, 3, 4, 5, 6, 7,14 and every 14 days later. Pharmacokinetic parameters are calculated bymodel independent methods on a Digital Equipment Corporation VAX 8600computer system using the latest version of the BIOAVL software. Thefollowing pharmacokinetics parameters are determined: peak serumconcentration (C_(max)); time to peak serum concentration (t_(max));area under the concentration-time curve (AUC) from time zero to the lastblood sampling time (AUC₀₋₇₂) calculated with the use of the lineartrapezoidal rule; and terminal elimination half-life (t_(1/2)), computedfrom the elimination rate constant. The elimination rate constant isestimated by linear regression of consecutive data points in theterminal linear region of the log-linear concentration-time plot. Themean, standard deviation (SD), and coefficient of variation (CV) of thepharmacokinetic parameters are calculated for each treatment.

Patient Response to Compound 4 Therapy:

Patient response is assessed via weekly hospital visits, weight andwaist measurements and routine blood tests.

Statistical Analysis:

The association of sodium taurocholate and risk of pancreatic cancer isanalyzed in multivariable logistic regression models including age, sex,race, smoking, alcohol, BMI, family history of cancer, duration ofdiabetes, and insulin use. To control for reversal causality due topancreatic cancer-caused diabetes, risk of pancreatic cancer isestimated after exclusion of those with duration of diabetes ≤2 years.HbA1c level (≤7% or >7%), a marker of glycemic control, is comparedbetween placebo and users of sodium taurocholate by ω2 test. Thedemographic and risk factors as well as duration of diabetes and insulinuse are compared between sodium taurocholate users and placebo by χ2test.

While preferred embodiments of the present invention have been shown anddescribed herein, it will be obvious to those skilled in the art thatsuch embodiments are provided by way of example only. Numerousvariations, changes, and substitutions will now occur to those skilledin the art without departing from the invention. It should be understoodthat various alternatives to the embodiments of the invention describedherein may be employed in practicing the invention. It is intended thatthe following claims define the scope of the invention and that methodsand structures within the scope of these claims and their equivalents becovered thereby.

What is claimed is:
 1. A method of increasing the concentration of bile acids and salts thereof in the distal gastrointestinal tract of an individual comprising administering an Apical Sodium-dependent Bile Acid Transporter Inhibitor (ASBTI) to the individual, wherein a therapeutically amount of the ASBTI is delivered to the distal ileum, the colon, or the rectum of an individual in need thereof, wherein the ASBTI is selected from

and potassium((2R,3R,4S,5R,6R)-4-benzyloxy-6-{3-[3-((3S,4R,5R)-3-butyl-7-dimethylamino-3-ethyl-4-hydroxy-1,1-dioxo-2,3,4,5-tetrahydro-1H-benzo[b]thiepin-5-yl)-phenyl]-ureido}-3,5-dihydroxy-tetrahydropyran-2-ylmethyl)sulphate ethanolate hydrate.
 2. The method of claim 1, further comprising administering a second agent selected from an enteroendocrine peptide enhancing agent, a nuclear farnesoid X receptor (FXR) agonist, a liver receptor homolog 1 (LRH-1), a DPP-IV inhibitor, a proton pump inhibitor, H2 antagonist, prokinetic agent, a biguanide, an incretin mimetic, a thiazolidinone, a mucoadhesive agent, and GLP-1 or an analog thereof.
 3. The method of claim 1, wherein the individual is a prematurely born infant, an enterally-fed infant, or a formula-fed infant.
 4. The method of claim 1, wherein the ASBTI reduces intraenterocyte bile acids or reduces necrosis and/or damage to ileal architecture in an individual in need thereof.
 5. The method of claim 2, wherein the enteroendocrine peptide enhancing agent is a bile acid, a bile salt, a bile acid mimic, a bile salt mimic, or a combination thereof, optionally wherein the bile acid or the bile acid mimic is cholic acid, deoxycholic acid, glycocholic acid, glycodeoxycholic acid, taurocholic acid, taurodihydrofusidate, taurodeoxycholic acid, cholate, glycocholate, deoxycholate, taurocholate, taurodeoxycholate, chenodeoxycholic acid, a TGR5-binding analog, M-BAR agonist, GPR119 agonist, GPR120 agonist, GPR131 agonist, GPR140 agonist, GPR143 agonist, GPBAR1 agonist, BG37 agonist, FXR agonist, 6-methyl-2-oxo-4-thiophen-2-yl-1,2,3,4-tetrahydro-pyrimidine-5-carboxylic acid benzyl ester, INT-777, RG-239, oleanolic acid, or crataegolic acid, or a compound represented by Formula (VII):

wherein: each R¹ is independently H, OH, lower alkyl, or lower heteroalkyl; L is a substituted or unsubstituted alkyl or substituted or unsubstituted heteroalkyl; each R² is independently H, OH, lower alkyl, or lower heteroalkyl; R³ is H, OH, O-lower alkyl, lower alkyl, or lower heteroalkyl; A is COOR⁴, S(O)_(n)R⁴, or OR⁵; R⁴ is H, an anion, a pharmaceutically acceptable cation, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or an amino acid; n is 1-3; R⁵ is lower alkyl or H; or a salt thereof.
 6. The method of claim 2, wherein the FXR agonist is GW4064, GW9662, INT-747, T0901317, WAY-362450, fexaramine, cholic acid, deoxycholic acid, glycocholic acid, glycodeoxycholic acid, taurocholic acid, taurodihydrofusidate, taurodeoxycholic acid, cholate, glycocholate, deoxycholate, taurocholate, taurodeoxycholate, chenodeoxycholic acid, or a salt thereof, or a combination thereof.
 7. The method of claim 2, wherein the ASBTI and/or the second agent is administered before ingestion of food.
 8. The method of claim 2, wherein the ASBTI and/or the second agent is administered less than about 60 minutes before ingestion of food.
 9. The method of claim 2, wherein the ASBTI and/or the second agent is administered less than about 30 minutes before ingestion of food.
 10. The method of claim 2, wherein the ASBTI and/or the second agent is administered orally.
 11. The method of claim 2, wherein the ASBTI and/or the second agent is administered orally as an ileal-pH sensitive release or an enterically coated formulation.
 12. The method of claim 2, wherein the ASBTI is


13. The method of claim 2, wherein the ASBTI is potassium((2R,3R,4S,5R,6R)-4-benzyloxy-6-{3-[3-((3S,4R,5R)-3-butyl-7-dimethylamino-3-ethyl-4-hydroxy-1,1-dioxo-2,3,4,5-tetrahydro-1H-benzo[b]thiepin-5-yl)-phenyl]-ureido}-3,5-dihydroxy-tetrahydropyran-2-ylmethyl)sulphate ethanolate hydrate, or a pharmaceutically acceptable salt or solvate thereof.
 14. The method of claim 1, wherein the subject does not have diabetes.
 15. The method of claim 1, wherein the ASBTI is administered before ingestion of food.
 16. The method of claim 1, wherein the ASBTI is administered less than about 60 minutes before ingestion of food.
 17. The method of claim 1, wherein the ASBTI is administered less than about 30 minutes before ingestion of food.
 18. The method of claim 1, wherein the ASBTI is administered orally.
 19. The method of claim 1, wherein the ASBTI is administered orally as an ileal-pH sensitive release or an enterically coated formulation.
 20. The method of claim 1, wherein the ASBTI is


21. The method of claim 1, wherein the ASBTI is potassium((2R,3R,4S,5R,6R)-4-benzyloxy-6-{3-[3-((3S,4R,5R)-3-butyl-7-dimethylamino-3-ethyl-4-hydroxy-1,1-dioxo-2,3,4,5-tetrahydro-1H-benzo[b]thiepin-5-yl)-phenyl]-ureido}-3,5-dihydroxy-tetrahydropyran-2-ylmethyl)sulphate ethanolate hydrate, or a pharmaceutically acceptable salt or solvate thereof. 